Skip Navigation

This Article
Right arrow Print PDF (7715K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Citing Articles
Right arrowScopus Links
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by La Branche, H.
Right arrow Articles by Chabot, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by La Branche, H.
Right arrow Articles by Chabot, B.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1991, Vol. 19, No. 16 4509-4514
© 1991

MOLECULAR BIOLOGY

Proteolysis of splicing factors during rat and monkey cell fractionation

Hélène La Branche, Danielle Frappier and Benoit Chabot*

Département de Microbiologie, Faculté de Médecine, Université de Sherbrooke Sherbrooke, Québec J1H 5N4, Canada

* To whom correspondence should be addressed

Received April 8, 1991. Revised July 19, 1991. Accepted July 19, 1991.

We have investigated the ability of various rat and monkey cell lines to yield nuclear extracts that would allow splicing of a model adenovirus pre-mRNA substrate. Extracts from normal FR3T3, rat-1 and CV-1 fibroblasts were unable to assemble splicing complexes and displayed a dramatic reduction in the binding activity of the splicing factor 65 kD U2AF. These results correlated with reduced levels of 65 kD U2AF and the snRNP-associated B protein. When a battery of protease inhibitors was used during cell fractionation, increased levels of 65 kD U2AF and B proteins were detected. Most importantly, U2AF binding and complex formation were dramatically improved in FR3T3, rat-1 and CV-1 extracts. Interestingly, transformation of rat and monkey cells with the SV40 large T antigen yielded extracts active in complex formation. Similar extracts were generated following transformation of rat-1 cells with the Py middle T antigen but not with the v-fos oncogene. Only SV40-transformed FR3T3 extracts displayed splicing activity. Our results indicate that proteolysis is a major obstacle encountered during the preparation of active extracts from normal rat and monkey cells and suggest that cells transformed with T antigens manifest reduced proteolysis during fractionation.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Mol. Biol. CellHome page
Y. Shav-Tal, M. Cohen, S. Lapter, B. Dye, J. G. Patton, J. Vandekerckhove, and D. Zipori
Nuclear Relocalization of the Pre-mRNA Splicing Factor PSF during Apoptosis Involves Hyperphosphorylation, Masking of Antigenic Epitopes, and Changes in Protein Interactions
Mol. Biol. Cell, August 1, 2001; 12(8): 2328 - 2340.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. L. Chew, L. Baginsky, and I. C. Eperon
An exonic splicing silencer in the testes-specific DNA ligase III {beta} exon
Nucleic Acids Res., January 15, 2000; 28(2): 402 - 410.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Côté, J. Beaudoin, R. Tacke, and B. Chabot
The U1 Small Nuclear Ribonucleoprotein/5` Splice Site Interaction Affects U2AF[IMAGE] Binding to the Downstream 3` Splice Site
J. Biol. Chem., February 24, 1995; 270(8): 4031 - 4036.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
P. Norton
Alternative pre-mRNA splicing: factors involved in splice site selection
J. Cell Sci., January 1, 1994; 107(1): 1 - 7.
[PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.