From owner-ageing@net.bio.net Mon Jan 03 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: Sydney Shall Newsgroups: bionet.molbio.ageing Subject: Werner's Syndrome and cell Ageing Date: 4 Jan 1994 13:19:42 -0000 Lines: 44 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2gbqde$bpc@mserv1.dl.ac.uk> Original-To: AGEING@dl.ac.uk Further to the posting from Jim Cummins this morning (?), I would add that several groups have now described the inhibitor of DNA replication that seems to operate in senescent cells. See the latest issue of Cell (December 1993), of Science and of Nature (at the end of 1993) and the first issue of Experimental cell Research, January 1994 (an article by Jim Smith, form Houston, USA). This is an important step I think in beginning to understand the mechanics used to prevent senescent cells from replicating. A Happy New Year to you all!!!!!!! ************************************************************************** ************************************************************************** Sydney SHALL, Laboratory of Cell and Molecular Biology, Biology Building, University of Sussex, Brighton, East Sussex BN1 9QG, ENGLAND. Telephone: +44.273.67.83.03 FAX: +44.273.67.84.33 E-Mail: Janet: S.Shall@uk.ac.sussex Elsewhere: S.Shall@sussex.ac.uk EARN/BITNET: S.Shall%sussex@ukacrl ******************************************************************************* ******************************************************************************* From owner-ageing@net.bio.net Tue Jan 04 22:00:00 1994 Path: biosci!daresbury!doc.ic.ac.uk!agate!howland.reston.ans.net!news.moneng.mei.com!uwm.edu!msuinfo!harbinger.cc.monash.edu.au!uniwa!newsman!Jim.Cummins From: cummins@possum.murdoch.edu.au (Jim Cummins) Newsgroups: bionet.molbio.ageing Subject: Re: Werner's Syndrome and cell Ageing Date: 5 Jan 1994 02:32:05 GMT Organization: Murdoch University, Western Australia Lines: 36 Sender: -Not-Authenticated-[2792] Message-ID: <2gd8r5INNag0@newsman.csu.murdoch.edu.au> References: <2gbqde$bpc@mserv1.dl.ac.uk> NNTP-Posting-Host: vetmac3.csu.murdoch.edu.au X-Posted-From: InterNews 1.0@newsman.csu.murdoch.edu.au. Xdisclaimer: No attempt was made to authenticate the sender's name. Is this anything to do with telomere shortenting with age: a thread that developed on this newsgroup a while back? The following is a copy of a posting from Dr Baranidharan, of Bsngalore Read Burnside,E. (1984) Ann.Rev.BioChem Vol.53, p163. (there is also a review article in Scientific American on this and I will try to fish it out if someone is interested) for the first discussion on telomeric repeats. The Telomere (TTAGGG)n repeats at both ends of chromosomes in all the vertebrates. The structure is antiparallel quadruplet and binds to an enzyme called 'telomerase' which is the first enzyme to be discovered that contains an RNA that is essential for its activity (this threw out the protein-enzyme hypothesis). This enzyme is found mostly in the germ cells (where spermatozoa production takes place) and NOT in somatic cells. It is also known that the TTAGGG repeats disable the shortening of chromosomes. However the cell ageing and deletions in the (TTAGGG)n form a chicken-and-egg problem. Others who are working on this problem include A.Rich (MIT). S.Baranidharan Molecular Biophysics Unit Indian Institute of Science Bangalore 560 012 India. barani@mbu.iisc.ernet.in Jim Cummins School of Veterinary Studies Murdoch University Western Australia 6150 Tel +61-9-360 2668 Fax +61-9-310 4144 For every complex problem there's a simple solution. And it's wrong! From owner-ageing@net.bio.net Wed Jan 05 22:00:00 1994 Path: biosci!daresbury!doc.ic.ac.uk!agate!howland.reston.ans.net!gatech!concert!news-feed-1.peachnet.edu!umn.edu!lynx.unm.edu!triton.unm.edu!news-user From: varela@unm.edu Newsgroups: bionet.molbio.ageing Subject: Can the age of a cell be ascertained? Date: 6 Jan 1994 07:30:07 -0700 Organization: University of New Mexico, Albuquerque Lines: 13 Message-ID: <2gh79f$hf9@triton.unm.edu> NNTP-Posting-Host: triton.unm.edu Hi, Someone in our laboratory posed the following question: If a cell is plucked from lets say the liver (or any other tissue for that matter), is there any way to ascertain how many times that cell has undergone cell division? We look forward to receiving your comments. Please E-mail either myself (mvarela@medusa.unm.edu) or Kent who posed the question (kent@triton.unm.edu). Thank you in advance!!! Manny Varela mvarela@medusa.unm.edu UNM SOM From owner-ageing@net.bio.net Mon Jan 10 22:00:00 1994 Path: biosci!bcm!cs.utexas.edu!sdd.hp.com!nigel.msen.com!yale.edu!yale!hsdndev!rascal.med.harvard.edu!RIYER From: riyer@rascal.med.harvard.edu Newsgroups: bionet.cellbiol,bionet.drosophila,bionet.general,bionet.molbio.ageing,bionet.molbio.genome-program,bionet.molbio.hiv,bionet.molbio.yeast,bionet.molbio.methds-reagnts,bionet.neuroscience,bionet.virology Subject: Reporter gene vectors Date: 11 Jan 1994 02:08:03 GMT Organization: Harvard Medical School, Department of Genetics Lines: 19 Message-ID: <2gt1m4$o1r@hsdndev.harvard.edu> Reply-To: riyer@rascal.med.harvard.edu NNTP-Posting-Host: rascal.med.harvard.edu Xref: biosci bionet.cellbiol:230 bionet.drosophila:202 bionet.general:7045 bionet.molbio.ageing:609 bionet.molbio.genome-program:613 bionet.molbio.hiv:281 bionet.molbio.yeast:593 bionet.molbio.methds-reagnts:10176 bionet.neuroscience:2307 bionet.virology:348 I recd. so many requests from people who did not get my earlier post that I decided to save time by re-posting it. At the outset I would like to state that I am only a satisfied user of the above vectors and am not associated with the company that markets them. pow Hi netters, I would like to report the availability of a set of reporter gene vectors that have some distinct advantages over those currently in use for the purpose of analysis of eukaryotic promoters & enhancers. The vectors , SV40-Syncat, Syncat I, Syncat II, SV40-pFlash, pFlash I and pFlash II, are as their names suggest CAT & Luciferase (Luc) reporter gene vectors. What distinguishes them from other vectors in this category is that these vectors produce near-zero background. This feature is due to the fact, that they use a modified SV40-t-intron as the donor for the poly-adenylation signal. The wild type SV40-t-intron is the generic donor for this function and is widely used in a variety of reporter gene vectors, eg. pCAT- basic, pCAT-promoter, pGL-basic and pGL-promoter (Promega). However one little known fact about the wild-type-t-intron is that it contains cryptic enhancer sequences that produce significant background activity, thereby raising the threshold of sensitivity. Until now this feature was not a serious limitation since scientist were analysing strong promoters/enhancers that produced high signal to noise ratios. However the focus in the past couple of years has shifted to the analysis of weak promoters, or promoters that have very low basal transcription rates but are specifically induced to high levels by cytokines, or promoters that function only in cells that are very poorly transfectable. Analysis of such regulatory elements requires systems of high sensitivity as wel l as specificity. The Syncat & pFlash series of vectors provide precisely these capabilities, since they contain a t-intron polyA signal that has been modified to be devoid of cryptic enhancer activity. The 2nd. feature of these vectors is the choice of the heterologous promoter in Syncat II and pFlash II. These vectors use the well characterized HSV-tk TATA box containing minimal promoter situated immediately upstream of the reporter gene. The HSV-tk is a widely used basal promoter that has been do cumented to produce no spurious interactions with enhancers of interest. By contrast vectors like pCAT-promoter or pGL-promoter use the SV40 promoter. The SV40 promoter contains an atypical TATA-box and more seriously has been documented to spuriously repress certain cytokine inducible enhancers (Benech, et.al.. J.Exp.Med. 1992, Vol. 176: 1115-1123). I should know, because I am one of the authors of that paper and this defect in the pCAT-promoter vector cost me 6 months of time and nearly resulted i n our being scooped by a competing lab. Other features of these vectors include a very versatile multiple cloning site upstream and downstream of the reporter gene cassette, flanked by T3 & T7 promoter sequences for direct sequencing and creation of unidirectionally deleted mutant libraries as well as f1 origin of replication for ssDNA recovery and site-directed mutagenesis. The combination of these features enabled me to perform all my DNA manipulations from cloning of a 5 kb genomic 5U flanking r egion upstream of the reporter gene ---to--- creation of nested deletion mutant libraries followed by sequence verification ---to--- ssDNA-site-directed mutagenesis and transfection of each construct --- ALL IN THE SAME REPORTER GENE VECTOR I STARTED WITH. The time savings alone were upto 50%. These vectors are available from SynapSys Corp. To get more info about them send E-mail to :- # # # # synapsys@world.std.com. # From owner-ageing@net.bio.net Mon Jan 10 22:00:00 1994 Newsgroups: bionet.cellbiol,bionet.drosophila,bionet.general,bionet.molbio.ageing,bionet.molbio.genome-program,bionet.molbio.hiv,bionet.molbio.yeast,bionet.molbio.methds-reagnts,bionet.neuroscience,bionet.virology Path: biosci!daresbury!doc.ic.ac.uk!uknet!pipex!howland.reston.ans.net!EU.net!ub4b!reks.uia.ac.be!news From: przemko@reks.uia.ac.be (Przemko) Subject: Re: Reporter gene vectors Message-ID: <1994Jan11.083738.26257@reks.uia.ac.be> Sender: news@reks.uia.ac.be (USENET News System) Organization: University of Antwerp X-Newsreader: Date: Tue, 11 Jan 1994 08:37:38 GMT Lines: 39 Xref: biosci bionet.cellbiol:231 bionet.drosophila:203 bionet.general:7051 bionet.molbio.ageing:610 bionet.molbio.genome-program:615 bionet.molbio.hiv:282 bionet.molbio.yeast:596 bionet.molbio.methds-reagnts:10183 bionet.neuroscience:2308 bionet.virology:349 In article <2gt1m4$o1r@hsdndev.harvard.edu> riyer@rascal.med.harvard.edu writes: >I recd. so many requests from people who did not get my earlier post that I >decided to save time by re-posting it. At the outset I would like to state >that I am only a satisfied user of the above vectors and am not associated with the company that markets them. > > >pow >Hi netters, I would like to report the availability of a set of reporter gene vectors that have some distinct advantages over those currently in use for the purpose of analysis of eukaryotic promoters & enhancers. The vectors , SV40-Syncat, Syncat I, Syncat II, SV40-pFlash, pFlash I and pFlash II, are as their names suggest CAT & Luciferase (Luc) reporter gene vectors. What distinguishes them from other vectors in this category is that these vectors produce near-zero background. This feature > > >to the fact, that they use a modified SV40-t-intron as the donor for the poly-adenylation signal. The wild type SV40-t-intron is the generic donor for this function and is widely used in a variety of reporter gene vectors, eg. pCAT- basic, pCAT-promoter, pGL-basic and pGL-promoter (Promega). However one little known fact about the wild-type-t-intron is that it contains cryptic enhancer sequences that produce significant background activity, thereby raising the threshold of sensitivity. Until n > >s > feature was not a serious limitation since scientist were analysing strong promoters/enhancers that produced high signal to noise ratios. However the focus in the past couple of years has shifted to the analysis of weak promoters, or promoters that have very low basal transcription rates but are specifically induced to high levels by cytokines, or promoters that function only in cells that are very poorly transfectable. Analysis of such regulatory elements requires systems of high sensitivity > >l >l as specificity. The Syncat & pFlash series of vectors provide precisely these capabilities, since they contain a t-intron polyA signal that has been modified to be devoid of cryptic enhancer activity. The 2nd. feature of these vectors is the choice of the heterologous promoter in Syncat II and pFlash II. These vectors use the well characterized HSV-tk TATA box containing minimal promoter situated immediately upstream of the reporter gene. The HSV-tk is a widely used basal promoter that has > >o >cumented to produce no spurious interactions with enhancers of interest. By contrast vectors like pCAT-promoter or pGL-promoter use the SV40 promoter. The SV40 promoter contains an atypical TATA-box and more seriously has been documented to spuriously repress certain cytokine inducible enhancers (Benech, et.al.. J.Exp.Med. 1992, Vol. 176: 1115-1123). I should know, because I am one of the authors of that paper and this defect in the pCAT-promoter vector cost me 6 months of time and nearly res > >i >n our being scooped by a competing lab. Other features of these vectors include a very versatile multiple cloning site upstream and downstream of the reporter gene cassette, flanked by T3 & T7 promoter sequences for direct sequencing and creation of unidirectionally deleted mutant libraries as well as f1 origin of replication for ssDNA recovery and site-directed mutagenesis. The combination of these features enabled me to perform all my DNA manipulations from cloning of a 5 kb genomic 5U fla > >r >egion upstream of the reporter gene ---to--- creation of nested deletion mutant libraries followed by sequence verification ---to--- ssDNA-site-directed mutagenesis and transfection of each construct --- ALL IN THE SAME REPORTER GENE VECTOR I STARTED WITH. The time savings alone were upto 50%. These vectors are available from SynapSys Corp. To get more info about them send E-mail to :- # ># ># ># >synapsys@world.std.com. # > > Yeah, say either my newsreader is #@$&^%@ or you AGAIN forgot CR after 80 characters. COULD YOU, PLEASE, REPOST IT IN A MORE READABLE FORM? Przemko From owner-ageing@net.bio.net Mon Jan 10 22:00:00 1994 Path: biosci!daresbury!doc.ic.ac.uk!uknet!pipex!howland.reston.ans.net!xlink.net!fauern!lrz-muenchen.de!ipp-garching.mpg.de!alf.biochem.mpg.de!krasel From: krasel@alf.biochem.mpg.de (Cornelius Krasel) Newsgroups: bionet.cellbiol,bionet.drosophila,bionet.general,bionet.molbio.ageing,bionet.molbio.genome-program,bionet.molbio.hiv,bionet.molbio.yeast,bionet.molbio.methds-reagnts,bionet.neuroscience,bionet.virology Subject: Re: Reporter gene vectors Followup-To: bionet.cellbiol,bionet.drosophila,bionet.general,bionet.molbio.ageing,bionet.molbio.genome-program,bionet.molbio.hiv,bionet.molbio.yeast,bionet.molbio.methds-reagnts,bionet.neuroscience,bionet.virology Date: 11 Jan 1994 11:58:10 GMT Organization: Rechenzentrum der Max-Planck-Gesellschaft in Garching Lines: 75 Message-ID: <2gu48iINN1apu@sat.ipp-garching.mpg.de> References: <2gt1m4$o1r@hsdndev.harvard.edu> NNTP-Posting-Host: alf.biochem.mpg.de X-Newsreader: TIN [version 1.1 PL8] Xref: biosci bionet.cellbiol:232 bionet.drosophila:204 bionet.general:7054 bionet.molbio.ageing:611 bionet.molbio.genome-program:616 bionet.molbio.hiv:283 bionet.molbio.yeast:598 bionet.molbio.methds-reagnts:10188 bionet.neuroscience:2309 bionet.virology:350 riyer@rascal.med.harvard.edu wrote: : I recd. so many requests from people who did not get my earlier post that I : decided to save time by re-posting it. At the outset I would like to state : that I am only a satisfied user of the above vectors and am not associated : with the company that markets them. And now here comes a readable version (with <80 chars/line) :-) : Hi netters, I would like to report the availability of a set of : reporter gene vectors that have some distinct advantages over those : currently in use for the purpose of analysis of eukaryotic promoters : & enhancers. The vectors , SV40-Syncat, Syncat I, Syncat II, : SV40-pFlash, pFlash I and pFlash II, are as their names suggest : CAT & Luciferase (Luc) reporter gene vectors. What distinguishes : them from other vectors in this category is that these vectors produce : near-zero background. This feature is due to the fact, that they use a : modified SV40-t-intron as the donor for the poly-adenylation signal. : The wild type SV40-t-intron is the generic donor for this function : and is widely used in a variety of reporter gene vectors, eg. pCAT- : basic, pCAT-promoter, pGL-basic and pGL-promoter (Promega). : However one little known fact about the wild-type-t-intron is that it : contains cryptic enhancer sequences that produce significant : background activity, thereby raising the threshold of sensitivity. : Until now this feature was not a serious limitation since scientist : were analysing strong promoters/enhancers that produced high : signal to noise ratios. However the focus in the past couple of : years has shifted to the analysis of weak promoters, or promoters : that have very low basal transcription rates but are specifically : induced to high levels by cytokines, or promoters that function only : in cells that are very poorly transfectable. Analysis of such : regulatory elements requires systems of high sensitivity as well as : specificity. The Syncat & pFlash series of vectors provide precisely : these capabilities, since they contain a t-intron polyA signal that has : been modified to be devoid of cryptic enhancer activity. : The 2nd. feature of these vectors is the choice of the : heterologous promoter in Syncat II and pFlash II. These vectors use : the well characterized HSV-tk TATA box containing minimal : promoter situated immediately upstream of the reporter gene. The : HSV-tk is a widely used basal promoter that has been documented : to produce no spurious interactions with enhancers of interest. By : contrast vectors like pCAT-promoter or pGL-promoter use the SV40 : promoter. The SV40 promoter contains an atypical TATA-box and : more seriously has been documented to spuriously repress certain : cytokine inducible enhancers (Benech, et.al.. J.Exp.Med. 1992, : Vol. 176: 1115-1123). I should know, because I am one of the : authors of that paper and this defect in the pCAT-promoter vector : cost me 6 months of time and nearly resulted in our being scooped : by a competing lab. Other features of these vectors include a very : versatile multiple cloning site upstream and downstream of the : reporter gene cassette, flanked by T3 & T7 promoter sequences for : direct sequencing and creation of unidirectionally deleted mutant : libraries as well as f1 origin of replication for ssDNA recovery and : site-directed mutagenesis. The combination of these features enabled : me to perform all my DNA manipulations from cloning of a 5 kb : genomic 5U flanking region upstream of the reporter gene ---to--- : creation of nested deletion mutant libraries followed by sequence : verification ---to--- ssDNA-site-directed mutagenesis and : transfection of each construct --- ALL IN THE SAME REPORTER : GENE VECTOR I STARTED WITH. The time savings alone were : upto 50%. These vectors are available from SynapSys Corp. To : get more info about them send E-mail to :- # : # : # : # : synapsys@world.std.com. # Just wanted to note that I am not affiliated with this company either. In fact, I have never worked with reporter genes :-) --Cornelius. -- /* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */ /* email: krasel@alf.biochem.mpg.de fax: +49 89 8578 3975 */ /* "People are DNA's way of making more DNA." (R. Dawkins/anonymous) */ From owner-ageing@net.bio.net Wed Jan 12 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!uchinews!ellis!bmdelane From: bmdelane@ellis.uchicago.edu (Brian Manning Delaney) Subject: Re: Werner's Syndrome and cell Ageing Message-ID: <1994Jan13.151255.9189@midway.uchicago.edu> Sender: news@uchinews.uchicago.edu (News System) Reply-To: bmdelane@midway.uchicago.edu Organization: University of Chicago References: <2gbqde$bpc@mserv1.dl.ac.uk> <2gd8r5INNag0@newsman.csu.murdoch.edu.au> Date: Thu, 13 Jan 1994 15:12:55 GMT Lines: 12 In article <2gd8r5INNag0@newsman.csu.murdoch.edu.au> cummins@possum.murdoch.edu.au (Jim Cummins) writes: >Is this anything to do with telomere shortenting with age: a thread >that developed on this newsgroup a while back? The following is a copy >of a posting from Dr Baranidharan, of Bsngalore > [....] > This enzyme [telomerase] is found >mostly in the germ cells (where spermatozoa production takes place) ^^^^^^ Where else is it found? -Brian Delaney From owner-ageing@net.bio.net Wed Jan 12 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!hubcap!darwin.sura.net!howland.reston.ans.net!torn!nott!cunews!superior!dsmillie From: dsmillie@superior.carleton.ca (David Smillie) Subject: Werner's Syndrome (reply) Message-ID: Sender: news@cunews.carleton.ca (News Administrator) Organization: Carleton University X-Newsreader: TIN [version 1.2 PL0] Date: Thu, 13 Jan 1994 19:33:06 GMT Lines: 18 Howdy, I used to be at McMaster University where Calvin Harley was doing his work into telomere shortening. One of his students showed that there was telomerase activity in immortal cell lines and cancer cells, but not in non-immortalized cell lines. Cal is currently working for some company in California, but I don't know how to reach him. Pretty neat results, actually. _____________________________________________________________________________ David Smillie, Carleton University Ancora imparo "I'd rather be happy than right any day." "But are you?" "What?" "Happy." "No," he said with a sigh, "That's where it all falls down, of course." _____________________________________________________________________________ From owner-ageing@net.bio.net Wed Jan 12 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: Sydney Shall Newsgroups: bionet.molbio.ageing Subject: Telomerase Date: 13 Jan 1994 20:12:09 -0000 Lines: 41 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2h49up$cgc@mserv1.dl.ac.uk> Original-To: AGEING@dl.ac.uk In answer to recent query as to where the enzyme telomerase is found; my understanding is that it is found in germ cells, but it is also found in cancer cell lines that grow indefinitely. This latter fact is part of the evidence that this enzyme is required for immortal growth, and that its absence is responsible for mortalization of a culture, becuase without this enzyme the telomeres in the cells will shorten till they finally reach a fatal threshold. I hope this helps. ************************************************************************** ************************************************************************** Sydney SHALL, Laboratory of Cell and Molecular Biology, Biology Building, University of Sussex, Brighton, East Sussex BN1 9QG, ENGLAND. Telephone: +44.273.67.83.03 FAX: +44.273.67.84.33 E-Mail: Janet: S.Shall@uk.ac.sussex Elsewhere: S.Shall@sussex.ac.uk EARN/BITNET: S.Shall%sussex@ukacrl ******************************************************************************* ******************************************************************************* From owner-ageing@net.bio.net Mon Jan 17 22:00:00 1994 Path: biosci!s.u-tokyo!kuis-news!wnoc-kyo-news!aist-nara!wnoc-tyo-news!news.u-tokyo.ac.jp!sinetnews!daffy!uwvax!uchinews!vixen.cso.uiuc.edu!howland.reston.ans.net!agate!headwall.Stanford.EDU!bcm!mbcr.bcm.tmc.edu!th035681 From: th035681@mbcr.bcm.tmc.edu (Tim Hughes) Newsgroups: bionet.molbio.ageing Subject: Something for the telomerase buffs Date: 17 Jan 1994 20:17:37 GMT Organization: Baylor College of Medicine, Houston, Tx Lines: 172 Sender: th035681@mbcr.bcm.tmc.edu (Timothy R. Hughes) Distribution: world Message-ID: <2herp1$r0o@gazette.bcm.tmc.edu> NNTP-Posting-Host: mbcr.bcm.tmc.edu I am a first year graduate student doing a rotation in Vicki Lundblad's lab. She discovered Est1, the yeast gene which holds the RNA template for the telomeres. It is a part of this "telomerase" complex. When it is disrupted, resulting yeast show a senescence phenotype mimicking eukaryotic senescence, but not exactly paralleling it - the yeasts die due to chromosome loss, whereas senescent fibroblasts can sit for years, and do not suffer massive chromosome loss or immediate death. I don't think they lose their telomeres, it's just that they stop dividing about the time their telomeres get real short. As far as I can tell, there is no way to tell if telomere shortening is a cause or effect of cell aging. For example, if the genome knows that humans don't live past a hundred years, it seems to me that it might just turn the telomerase off in somatic cells to save energy. It makes sense that cancer cells have turned it back on, because they are planning on dividing many more times than they're supposed to, since it has been shown in yeast that telomere loss leads to chromosome loss, which is definitely bad news for the cell. I think the big theory goiing around is that mammalian cells are supposed to sense the short telomeres, and exit the cell cycle, so they don't lose chromosomes perhaps. Cancer cells will die if they lose their chromosomes, so they have to turn the telomerase on. This can't be proven, though, or it would be on the cover of Time and Newsweek. In any case, I don't think it is yet possible to say whether short telomeres are an upstream or parallel occurrence with respect to cell senescence. When the genes involved in both processes are isolated and characterized it will be possible to say for certain. (I know I'm doing my part!) I pulled up a few abstracts on medline addressing questions posed in previous postings. It seems like the Canadian guys are really sold on this. AN 93066190. 93021. AU Allsopp-R-C. Vaziri-H. Patterson-C. Goldstein-S. Younglai-E-V. Futcher-A-B. Greider-C-W. Harley-C-B. IN Department of Biochemistry, McMaster University, Hamilton, ON, Canada. TI Telomere length predicts replicative capacity of human fibroblasts. SO Proc-Natl-Acad-Sci-U-S-A. 1992 Nov 1. 89(21). P 10114-8. JT PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. PT JOURNAL-ARTICLE (ART). AB When human fibroblasts from different donors are grown in vitro, only a small fraction of the variation in their finite replicative capacity is explained by the chronological age of the donor. Because we had previously shown that telomeres, the terminal guanine-rich sequences of chromosomes, shorten throughout the life-span of cultured cells, we wished to determine whether variation in initial telomere length would account for the unexplained variation in replicative capacity. Analysis of cells from 31 donors (aged 0-93 yr) indicated relatively weak correlations between proliferative ability and donor age (m = -0.2 doubling per yr; r = -0.42; P = 0.02) and between telomeric DNA and donor age (m = -15 base pairs per yr; r = -0.43; P = 0.02). However, there was a striking correlation, valid over the entire age range of the donors, between replicative capacity and initial telomere length (m = 10 doublings per kilobase pair; r = 0.76; P = 0.004), indicating that cell strains with shorter telomeres underwent significantly fewer doublings than those with longer telomeres. These observations suggest that telomere length is a biomarker of somatic cell aging in humans and are consistent with a causal role for telomere loss in this process. We also found that fibroblasts from Hutchinson-Gilford progeria donors had short telomeres, consistent with their reduced division potential in vitro. In contrast, telomeres from sperm DNA did not decrease with age of the donor, suggesting that a mechanism for maintaining telomere length, such as *telomerase* expression, may be active in germ-line tissue. Author-abstract. AN 92309418. 92000. AU Levy-M-Z. Allsopp-R-C. Futcher-A-B. Greider-C-W. Harley-C-B. IN Department of Biochemistry, McMaster University Hamilton, Ontario, Canada. TI Telomere end-replication problem and cell aging. SO J-Mol-Biol. 1992 Jun 20. 225(4). P 951-60. JT JOURNAL OF MOLECULAR BIOLOGY. PT JOURNAL-ARTICLE (ART). AB Since DNA polymerase requires a labile primer to initiate unidirectional 5'-3' synthesis, some bases at the 3' end of each template strand are not copied unless special mechanisms bypass this "end-replication" problem. Immortal eukaryotic cells, including transformed human cells, apparently use *telomerase,* an enzyme that elongates telomeres, to overcome incomplete end-replication. However, *telomerase* has not been detected in normal somatic cells, and these cells lose telomeres with age. Therefore, to better understand the consequences of incomplete replication, we modeled this process for a population of dividing cells. The analysis suggests four things. First, if single-stranded overhangs generated by incomplete replication are not degraded, then mean telomere length decreases by 0.25 of a deletion event per generation. If overhangs are degraded, the rate doubles. Data showing a decrease of about 50 base-pairs per generation in fibroblasts suggest that a full deletion event is 100 to 200 base-pairs. Second, if cells senesce after 80 doublings in vitro, mean telomere length decreases about 4000 base-pairs, but one or more telomeres in each cell will lose significantly more telomeric DNA. A checkpoint for regulation of cell growth may be signalled at that point. Third, variation in telomere length predicted by the model is consistent with the abrupt decline in dividing cells at senescence. Finally, variation in length of terminal restriction fragments is not fully explained by incomplete replication, suggesting significant interchromosomal variation in the length of telomeric or subtelomeric repeats. This analysis, together with assumptions allowing dominance of *telomerase* inactivation, suggests that telomere loss could explain cell cycle exit in human fibroblasts. Author-abstract. AN 91054430. 91000. AU Greider-C-W. IN Cold Spring Harbor Laboratory, New York 11724. TI Telomeres, *telomerase* and senescence. SO Bioessays. 1990 Aug. 12(8). P 363-9. JT BIOESSAYS. PT JOURNAL-ARTICLE (ART). REVIEW (REV). REVIEW-TUTORIAL (TUT). AB Eukaryotic chromosomes end with tandem repeats of simple sequences. These GC rich repeats allow telomere replication and stabilize chromosome ends. Telomere replication involves an equilibrium of sequence loss and addition at the ends of chromosomes. Repeats are added de novo by *telomerase,* an unusual DNA polymerase. *Telomerase* is an RNP in which an essential RNA component provides the template for the added telomere repeats. Telomere length maintenance plays an essential role in cell viability. Author-abstract. 58 Refs. AN 90259099. 90000. AU Harley-C-B. Futcher-A-B. Greider-C-W. IN Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada. TI Telomeres shorten during ageing of human fibroblasts. SO Nature. 1990 May 31. 345(6274). P 458-60. JT NATURE. PT JOURNAL-ARTICLE (ART). AB The terminus of a DNA helix has been called its Achilles' heel. Thus to prevent possible incomplete replication and instability of the termini of linear DNA, eukaryotic chromosomes end in characteristic repetitive DNA sequences within specialized structures called telomeres. In immortal cells, loss of telomeric DNA due to degradation or incomplete replication is apparently balanced by telomere elongation, which may involve de novo synthesis of additional repeats by novel DNA polymerase called *telomerase.* Such a polymerase has been recently detected in HeLa cells. It has been proposed that the finite doubling capacity of normal mammalian cells is due to a loss of telomeric DNA and eventual deletion of essential sequences. In yeast, the est1 mutation causes gradual loss of telomeric DNA and eventual cell death mimicking senescence in higher eukaryotic cells. Here, we show that the amount and length of telomeric DNA in human fibroblasts does in fact decrease as a function of serial passage during ageing in vitro and possibly in vivo. It is not known whether this loss of DNA has a causal role in senescence. Author-abstract. From owner-ageing@net.bio.net Mon Jan 17 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!s.u-tokyo!news.tisn.ad.jp!news.u-tokyo.ac.jp!sinetnews!daffy!uwvax!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!pipex!uknet!EU.net!ub4b!info-sparc1.info.ucl.ac.be!NewsWatcher!user From: klosen@bani.ucl.ac.be (Klosen Paul) Subject: Ageing of the cytoskeleton Message-ID: Followup-To: bionet.molbio.ageing Sender: news@info.ucl.ac.be (News Administrator) Nntp-Posting-Host: 130.104.130.2 Organization: Catholic University of Louvain Date: Tue, 18 Jan 1994 15:29:56 GMT Lines: 8 Can anyone provide me with the reference for a good review on the ageing of the cytoskeleton??? Thank You -- Paul Klosen Cell Biology Lab., Catholic University of Louvain klosen@bani.ucl.ac.be From owner-ageing@net.bio.net Tue Jan 18 22:00:00 1994 Path: biosci!CS.Arizona.EDU!math.arizona.edu!news.Arizona.EDU!hamblin.math.byu.edu!news.byu.edu!gatech!usenet.ins.cwru.edu!howland.reston.ans.net!cs.utexas.edu!uunet!munnari.oz.au!uniwa!newsman!Jim.Cummins From: cummins@possum.murdoch.edu.au (Jim Cummins) Newsgroups: bionet.molbio.ageing Subject: Re: Ageing of the cytoskeleton Date: 19 Jan 1994 05:10:53 GMT Organization: Murdoch University, Western Australia Lines: 16 Sender: -Not-Authenticated-[2792] Message-ID: <2hifctINNbl3@newsman.csu.murdoch.edu.au> References: NNTP-Posting-Host: vetmac3.csu.murdoch.edu.au X-Posted-From: InterNews 1.0@newsman.csu.murdoch.edu.au. Xdisclaimer: No attempt was made to authenticate the sender's name. In article klosen@bani.ucl.ac.be (Klosen Paul) writes: > Can anyone provide me with the reference for a good review on the ageing of > the cytoskeleton??? > Thank You Try Finch, CE; Longevity, senescence and the genome; U Chicago Press; 1990. This is pretty exhaustive. Unfortunately my grad student has stolen (borrowed) my copy so I can't check up on the cytoskeleton. Jim Cummins School of Veterinary Studies Murdoch University Western Australia 6150 Tel +61-9-360 2668 Fax +61-9-310 4144 "An inordinate fondness for beetles" From owner-ageing@net.bio.net Tue Jan 18 22:00:00 1994 Path: biosci!CS.Arizona.EDU!math.arizona.edu!news.Arizona.EDU!hamblin.math.byu.edu!sol.ctr.columbia.edu!howland.reston.ans.net!cs.utexas.edu!uunet!munnari.oz.au!uniwa!newsman!Jim.Cummins From: cummins@possum.murdoch.edu.au (Jim Cummins) Newsgroups: bionet.molbio.ageing Subject: Maternal effects on lifespan Date: 19 Jan 1994 07:31:35 GMT Organization: Murdoch University, Western Australia Lines: 21 Sender: -Not-Authenticated-[2792] Message-ID: <2hinknINNbvf@newsman.csu.murdoch.edu.au> NNTP-Posting-Host: vetmac3.csu.murdoch.edu.au X-Posted-From: InterNews 1.0@newsman.csu.murdoch.edu.au. Xdisclaimer: No attempt was made to authenticate the sender's name. A recent paper speculates that aspects of lifespan may be maternally inherited and mediated through mitochondrial DNA (Sont & Vandenbroucke, J Clin Epidemiol 46: 199-201, 1993). There is cerrtainly a strong association between age of death of offspring and mothers, but not fathers ( Abbott et al Am J Med Genet 2: 105-120, 1978) that has recently been confirmed in the Framingham longevity study (Brand et al J Clin Epidemiol 45: 169-174, 1992). We've now shown a relationship between mitochondrial DNA deletions and some forms of male infertility . Does anyone on the 'net know of any data that relates male infertility to (a) maternal age at conception (b) life expectancy (c) other forms of mitochondrial genome deficiency such as muscle weakness and dementia? Serious stuff aside, I guess the bottom line is that if you want to live long then pick your mother with care ;-) Jim Cummins School of Veterinary Studies Murdoch University Western Australia 6150 Tel +61-9-360 2668 Fax +61-9-310 4144 "An inordinate fondness for beetles"! From owner-ageing@net.bio.net Tue Jan 18 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: Sydney Shall Newsgroups: bionet.molbio.ageing Subject: Maternal effects on life-span. Date: 19 Jan 1994 10:09:44 -0000 Lines: 35 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2hj0t8$ddp@mserv1.dl.ac.uk> Original-To: AGEING@dl.ac.uk Further to the comment by Jim Cummins regarding maternal effects on life-span, I would draw attention to the well known phenomenon of maternal imprinting, which might be effected, in part, by methylation of DNA in the nucleus. ************************************************************************** ************************************************************************** Sydney SHALL, Laboratory of Cell and Molecular Biology, Biology Building, University of Sussex, Brighton, East Sussex BN1 9QG, ENGLAND. Telephone: +44.273.67.83.03 FAX: +44.273.67.84.33 E-Mail: Janet: S.Shall@uk.ac.sussex Elsewhere: S.Shall@sussex.ac.uk EARN/BITNET: S.Shall%sussex@ukacrl ******************************************************************************* ******************************************************************************* From owner-ageing@net.bio.net Tue Jan 18 22:00:00 1994 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!pipex!uknet!daresbury!not-for-mail From: Potter Wickware Newsgroups: bionet.molbio.ageing Subject: Re: Ageing of the cytoskeleton Date: 19 Jan 1994 19:34:16 -0000 Lines: 26 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2hk1vo$7k9@mserv1.dl.ac.uk> Original-To: Jim Cummins Not a review, but worht looking into: reports on the action of calpain against actin-binding protein. Yano et al, Thrombosis Research 71(5):385-96; 1993. Calpain of course being the "destroying angel" instrumental in the death of cells. On 19 Jan 1994, Jim Cummins wrote: > In article > klosen@bani.ucl.ac.be (Klosen Paul) writes: > > > Can anyone provide me with the reference for a good review on the ageing of > > the cytoskeleton??? > > Thank You > Try > Finch, CE; Longevity, senescence and the genome; U Chicago Press; 1990. > This is pretty exhaustive. Unfortunately my grad student has stolen > (borrowed) my copy so I can't check up on the cytoskeleton. > > Jim Cummins > School of Veterinary Studies > Murdoch University > Western Australia 6150 Tel +61-9-360 2668 Fax +61-9-310 4144 > "An inordinate fondness for beetles" > > From owner-ageing@net.bio.net Wed Jan 19 22:00:00 1994 Path: biosci!CS.Arizona.EDU!organpipe.uug.arizona.edu!math.arizona.edu!news.Arizona.EDU!hamblin.math.byu.edu!sol.ctr.columbia.edu!howland.reston.ans.net!agate!msuinfo!harbinger.cc.monash.edu.au!uniwa!newsman!Jim.Cummins From: cummins@possum.murdoch.edu.au (Jim Cummins) Newsgroups: bionet.molbio.ageing Subject: Re: Maternal effects on lifespan Date: 20 Jan 1994 01:14:01 GMT Organization: Murdoch University, Western Australia Lines: 20 Sender: -Not-Authenticated-[2792] Message-ID: <2hklspINNe50@newsman.csu.murdoch.edu.au> References: <2hinknINNbvf@newsman.csu.murdoch.edu.au> NNTP-Posting-Host: vetmac3.csu.murdoch.edu.au X-Posted-From: InterNews 1.0@newsman.csu.murdoch.edu.au. Xdisclaimer: No attempt was made to authenticate the sender's name. Sorry to be a bore on this topic, but it occurred to me last night that if life span is indeed maternally inherited in part, then there may be an association between maternal age at conception and the life expectancy of offspring. The thinking behind this is related to female meiotic pattern in which oocytes proliferate prenataslly and then enter a prolonged resting stage of M1, not reactivating until just before ovulation. There are well documented effects of maternal age on oocyte fertility and on chromosomal defects such as Downs syndrome. Aging oocytes might well acccumulate increased levels of defective mtDNA. While it would be very difficult to sort out lifestyle factors, this hypothesis would predict that in general younger siblings would live longer than older. Anyone in the group know of data that evaluates life expectancy in terms of birth order? Wonderful research area for a dedicated nutty epidemiologist! Jim Cummins School of Veterinary Studies Murdoch University Western Australia 6150 Tel +61-9-360 2668 Fax +61-9-310 4144 For every complex problem there's a simple solution. And it's wrong! From owner-ageing@net.bio.net Thu Jan 20 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!daresbury!doc.ic.ac.uk!warwick!uknet!nessie!ak02.scs.man.ac.uk!user From: a-kowald@nimr.mrc.ac.uk (Axel Kowald) Subject: Re: Maternal effects on lifespan Message-ID: Followup-To: bionet.molbio.ageing Sender: news@nessie.mcc.ac.uk (Usenet News System) Organization: Manchester Computing Centre References: <2hinknINNbvf@newsman.csu.murdoch.edu.au> <2hklspINNe50@newsman.csu.murdoch.edu.au> Date: Fri, 21 Jan 1994 12:25:50 GMT Lines: 25 In article <2hklspINNe50@newsman.csu.murdoch.edu.au>, cummins@possum.murdoch.edu.au (Jim Cummins) wrote: > Sorry to be a bore on this topic, but it occurred to me last night > that if life span is indeed maternally inherited in part, then there > may be an association between maternal age at conception and the life > expectancy of offspring. The thinking behind this is related to female Well, I don't know about those effects for humans, but for yeast a similar effect exists. Egilmez & Jazwinski (J. Bact., 171,37-42, 1989) showed that the generation times (cell doubling times) increased for old yeast cells. The interesting bit is that daughters from old mother cells had initially the same long generation times as the mother. However, after 3-4 divisions the generation times of the daughters decreased to the normal generation time for young cells. So it seems, there is some kind of maternally inherited effect. Egilmez & Jazwinski suggest some kind of substance which gets transmitted from mother to daughter and is then either diluted or degraded. Axel Kowald School of Biological Sciences University of Manchester England Tel +44-61-2755652 Fax +44-61-2755654 From owner-ageing@net.bio.net Sat Jan 22 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: Mr Georges L Friedli Newsgroups: bionet.molbio.ageing Subject: subscription Date: 23 Jan 1994 19:16:57 -0000 Lines: 2 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2huif9$p4m@mserv1.dl.ac.uk> Original-To: ageing@dl.ac.uk subscription for bsp1gf@surrey.ac.uk From owner-ageing@net.bio.net Sat Jan 22 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: caramelp@ERE.UMontreal.CA (Caramelli Paulo) Newsgroups: bionet.molbio.ageing Subject: Subscription Date: 23 Jan 1994 19:38:59 -0000 Lines: 3 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2hujoj$qdk@mserv1.dl.ac.uk> Original-To: AGEING@dl.ac.uk SUBSCRIBE AGEING PAULO CARAMELLI From owner-ageing@net.bio.net Sun Jan 23 22:00:00 1994 Path: biosci!daresbury!not-for-mail From: kapalla@devin.fns.uniba.sk (Marko Kapalla) Newsgroups: bionet.molbio.ageing Subject: Magnetic Field and Ageing Date: 24 Jan 1994 16:31:55 -0000 Lines: 34 Sender: daemon@mserv1.dl.ac.uk Distribution: bionet Message-ID: <2i0t5r$j1b@mserv1.dl.ac.uk> X-Char-Esc: 29 X-Charset: ASCII Original-To: ageing@dl.ac.uk (mailing list) Hallo everyone, Did any of you consider an influence of magnetic field (mg.field) of different intensities on ageing or lifespan, respectively? It is clear that the cell or the whole organism must respond to such a powerful enviromental factor as mg.field for sure is. Many of you might know about that many times denied Philadelphia Experiment, performed during the 2nd WW. Proceeding form Albert Einstein's theory of unitar field, they have tried to make a warship invisible using mg. field of enormous intensity. What is important of this experiment, is the strong influence of mg.fileld on all the living forms aboard including soliders, barins of who were terribly affected. On the other hand if the organism stays for some period in the no mg.field conditions it dies due to collapse of its biorhytms. And right this biorhytms, in my opinion, might be one of the keys to ageing. So, if we could control them through mg.field we might be able to control an ageing on a little bit higher level then the molecular one. Please, let me know how do you see the above mentioned, and whether there is the laboratory somewhere on this planet where they are interested in all this (if it is not TOP SECRET, of course). Thank you very much for paying attention Marko KAPALLA, currently at the 5th year of my study Dept of Biochemistry Mlynska Dolinna, CH-1 Comenius University, 842 15 Bratislava SLOVAKIA P.S. I relize that scientist should avoid refering to uncertain sources of information (Philadelphia Experiment) but anyway, somtimes, it can be helpful. From owner-ageing@net.bio.net Tue Jan 25 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!CS.Arizona.EDU!math.arizona.edu!news.Arizona.EDU!hamblin.math.byu.edu!sol.ctr.columbia.edu!howland.reston.ans.net!news.ans.net!malgudi.oar.net!sun!oucsace!ouvaxa.cats.ohiou.edu!JANAPATI From: janapati@ouvaxa.cats.ohiou.edu Subject: c-fos,c-myc probes Message-ID: Sender: usenet@oucsace.cs.ohiou.edu (Network News Poster) Reply-To: janapati@ouvaxa.cats.ohiou.edu Organization: Ohio University Computing & Technology Services Date: Tue, 25 Jan 1994 16:09:17 GMT Lines: 16 of these probes please reply. It will be of great help to me. I am working on ageing in rat brain cells. The probes should be preferably of rat sequences. I need c-fo of these probes please reply. It will be of great help to me. I need c-fos c-myc bcl-2 my address V. Janapati Janapati@ouvaxa.cats.ohiou.edu 310 irvine Hall Ohio university Athens, OH- 45701 From owner-ageing@net.bio.net Fri Jan 28 22:00:00 1994 Path: biosci!rutgers!biosci!CS.Arizona.EDU!math.arizona.edu!news.Arizona.EDU!hamblin.math.byu.edu!sol.ctr.columbia.edu!howland.reston.ans.net!news.ans.net!malgudi.oar.net!sun!oucsace!ouvaxa.cats.ohiou.edu!JANAPATI From: janapati@ouvaxa.cats.ohiou.edu Newsgroups: bionet.molbio.ageing Subject: c-fos,c-myc probes Message-ID: Date: 25 Jan 94 16:09:17 GMT Sender: usenet@oucsace.cs.ohiou.edu (Network News Poster) Reply-To: janapati@ouvaxa.cats.ohiou.edu Organization: Ohio University Computing & Technology Services Lines: 16 of these probes please reply. It will be of great help to me. I am working on ageing in rat brain cells. The probes should be preferably of rat sequences. I need c-fo of these probes please reply. It will be of great help to me. I need c-fos c-myc bcl-2 my address V. Janapati Janapati@ouvaxa.cats.ohiou.edu 310 irvine Hall Ohio university Athens, OH- 45701 From owner-ageing@net.bio.net Fri Jan 28 22:00:00 1994 Newsgroups: bionet.molbio.ageing Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!news.ans.net!malgudi.oar.net!sun!oucsace!ouvaxa.cats.ohiou.edu!JANAPATI From: janapati@ouvaxa.cats.ohiou.edu Subject: c-fos,c-myc probes Message-ID: Sender: usenet@oucsace.cs.ohiou.edu (Network News Poster) Reply-To: janapati@ouvaxa.cats.ohiou.edu Organization: Ohio University Computing & Technology Services Date: Tue, 25 Jan 1994 16:09:17 GMT Lines: 16 of these probes please reply. It will be of great help to me. I am working on ageing in rat brain cells. The probes should be preferably of rat sequences. I need c-fo of these probes please reply. It will be of great help to me. I need c-fos c-myc bcl-2 my address V. Janapati Janapati@ouvaxa.cats.ohiou.edu 310 irvine Hall Ohio university Athens, OH- 45701 From owner-ageing@net.bio.net Sat Jan 29 22:00:00 1994 Path: biosci!rutgers!cs.utexas.edu!howland.reston.ans.net!pipex!uunet!world!Oceania From: Oceania@world.std.com (Eric S Klien) Newsgroups: bionet.molbio.ageing Subject: Oceania Message-ID: Date: 30 Jan 94 02:17:02 GMT Organization: The World Public Access UNIX, Brookline, MA Lines: 11 Tired of the FDA's continous attacks on medical progress? On longer and longer waits for drug approval? On longer and longer waits for approval of new medical devices? On denials for new drugs despite evidence that the drugs benefits outweigh its side effects? Then learn about the new country Oceania where the government is prohibited from entering the health care system. Where you can take whatever therapy you wish or try any cure. To receive more information by e-mail, send your e-mail address to oceania@world.std.com.