IUBio

Weird protein behavior in cells - help.

David N. Levy levy at uab.edu
Wed Jan 6 15:24:23 EST 1999


I am studying a putative viral protein, whose behavior I have described
once before in this group, and received some useful responses.  I have
a new behavior I would like to describe which is quite baffling to me,
to see if anyone has a clue about it.  I sure don't.  I have expressed
this protein as a fusion product with GFP.  As I described in a
previous post, this protein is then seen to be either in the cytoplasm
or the nucleus, one or the other exclusively, in most cells.

Now I have made a fusion protein of just the first 43 amino acids (of
78) of my putative protein to GFP.  This protein is now found in the
cytoplasm, in a perinuclear pattern early (14 hours) after
transfection, then the interesting thing happens:  The cells become
bags of glowing balls.  Each cell is now comprised of about 50 to 100
round glowing vessicles.  In fact the whole cytoplasmic architecture is
gone and only these round vessicles are left.

I observed the same phenomenon by fusing the entire putative gene from
a divergent strain of virus to GFP, but I only saw this phenomenon when
I accidentally lysed the cells (unfixed) by lifting up the coverglass,
thus causing shear forces to disturb the cells.

I am posting a photo of this phenomenon on my computer at:

ftp://138.26.49.17/Spelunking%20Homunculus/Public/Viral%20protein%20phot
os/HeLa%20%26%20fusion%20protein.jpg

You can use Netscape to see this photo or download it and view it in
Photoshop or maybe other viewers (I don't know.)

Thanks.



David N. Levy
University of Alabama at Birmingham
Birmingham, AL 35294-0007

levy at uab.edu



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