SUMO interaction in a Yeast Two Hybrid Assay

redeamer alion85 at
Sat Feb 21 12:45:55 EST 2004

aji23 at (Andrew) wrote in message news:<958c94ef.0402200923.250d815c at>...
> Not sure how many of you readers work with SUMO, but I figured I'd
> post this notion that has been bothering me for a while now...

have a bit of experience with that.. from chromatin crosslinking and
immunoprecipitation :) know that SUMO is being regulated by Androgen

> In my reading I've come across many examples of people utilizing the
> Y2H assay to demonstrate interaction between their protein and SUMO. 
> Some investigators conclude based on their interaction data that this
> is SUMO conjugation; however, it is my understanding that Y2H shows
> noncovalent protein-protein interactions.  Since there is a clear
> difference between a protein's ability to physically interact with
> SUMO in a noncovalent fashion vs. its ability to be SUMO modified, is
> anyone else concerned with the interpretation of this data?

well, you are bothered about??? wheter your protein is covalently
linked to SUMO or noncovalently, as i understand? If so then that is
easy to control (at least theoretically):

do a chromatin crosslinking (formaldehyde, see one of my previous
posts on that)

1) yeasts without formaldehyde treatment

2) yeast with ......

then do an immunoprecipitation

analyze by western blot

was that your point?

o' zee bezt
-----------> Drew

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