|
|
|
|||||||
|
|||||||||
(Received for publication, May 1, 1996, and in revised form, August 12, 1996)
From the Division of Biology, California Institute of Technology,
Pasadena, California 91125
Pre-mRNA splicing takes place on a large
ribonucleoprotein particle, the spliceosome which contains the five
small nuclear ribonucleoproteins (snRNPs), U1, U2, U4, U5, and U6. In
Saccharomyces cerevisiae the mRNA splicing factors,
Prp9, Prp11, and Prp21, are necessary for addition of the U2 snRNP to
the pre-mRNA in an early step of spliceosome assembly. This paper
describes a study of interactions between these proteins and their role
in spliceosome assembly. The proteins were expressed in
Escherichia coli. Prp9 and Prp11 were purified by metal
affinity chromatography. Prp21 was purified using a
solubilization/renaturation protocol. We have combined these separately
purified proteins and present direct evidence of a Prp9·Prp11·Prp21
protein complex that is functional in in vitro
splicing assays. Characteristics of this Prp9·Prp11·Prp21 complex
were further investigated using proteins synthesized in
vitro. In addition, we found that Prp9, Prp11, and Prp21
influence the structure of the U2 snRNP in a manner that alters the
accessibility of the branch point pairing region of the U2 snRNA to
oligonucleotide- directed RNaseH cleavage. We present a model,
based on the data presented here and in the accompanying paper, for a
combined role of Prp9, Prp11, Prp21, and Prp5 in activating the U2
snRNP for assembly into the pre-spliceosome.
Volume 271, Number 52,
Issue of December 27, 1996
pp. 33268-33276
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  T. R. Kosowski, H. R. Keys, T. K. Quan, and S. W. Ruby DExD/H-box Prp5 protein is in the spliceosome during most of the splicing cycle RNA, July 1, 2009; 15(7): 1345 - 1362. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Qiu, J. Eifert, L. Wacheul, M. Thiry, A. C. Berger, J. Jakovljevic, J. L. Woolford Jr., A. H. Corbett, D. L. J. Lafontaine, R. M. Terns, et al. Identification of Genes That Function in the Biogenesis and Localization of Small Nucleolar RNAs in Saccharomyces cerevisiae Mol. Cell. Biol., June 1, 2008; 28(11): 3686 - 3699. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. J. Perriman and M. Ares Jr. Rearrangement of competing U2 RNA helices within the spliceosome promotes multiple steps in splicing Genes & Dev., April 1, 2007; 21(7): 811 - 820. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Perriman, I. Barta, G. K. Voeltz, J. Abelson, and M. Ares Jr. ATP requirement for Prp5p function is determined by Cus2p and the structure of U2 small nuclear RNA PNAS, November 25, 2003; 100(24): 13857 - 13862. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X.-h. Liang, A. Haritan, S. Uliel, and S. Michaeli trans and cis Splicing in Trypanosomatids: Mechanism, Factors, and Regulation Eukaryot. Cell, October 1, 2003; 2(5): 830 - 840. [Full Text] [PDF]
|
|
|
|
 |
|
 B. K. Abu Dayyeh, T. K. Quan, M. Castro, and S. W. Ruby Probing Interactions between the U2 Small Nuclear Ribonucleoprotein and the DEAD-box Protein, Prp5 J. Biol. Chem., June 7, 2002; 277(23): 20221 - 20233. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. J. Nixon, A. Wang, H. G. Morrison, A. G. McArthur, M. L. Sogin, B. J. Loftus, and J. Samuelson From the Cover: A spliceosomal intron in Giardialamblia PNAS, March 19, 2002; 99(6): 3701 - 3705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Nesic and A. Kramer Domains in Human Splicing Factors SF3a60 and SF3a66 Required for Binding to SF3a120, Assembly of the 17S U2 snRNP, and Prespliceosome Formation Mol. Cell. Biol., October 1, 2001; 21(19): 6406 - 6417. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Zhang and M. R. Green Identification and characterization of yUAP/Sub2p, a yeast homolog of the essential human pre-mRNA splicing factor hUAP56 Genes & Dev., January 1, 2001; 15(1): 30 - 35. [Abstract] [Full Text]
|
|
|
|
|
|
D. Libri, N. Graziani, C. Saguez, and J. Boulay Multiple roles for the yeast SUB2/yUAP56 gene in splicing Genes & Dev., January 1, 2001; 15(1): 36 - 41. [Abstract] [Full Text]
|
|
|
|
|
|
A. L. Kistler and C. Guthrie Deletion of MUD2, the yeast homolog of U2AF65, can bypass the requirement for Sub2, an essential spliceosomal ATPase Genes & Dev., January 1, 2001; 15(1): 42 - 49. [Abstract] [Full Text]
|
|
|
|
|
|
R. Perriman and M. Ares Jr. ATP can be dispensable for prespliceosome formation in yeast Genes & Dev., January 1, 2000; 14(1): 97 - 107. [Abstract] [Full Text]
|
|
|
|
|
|
E. J. Chen, A. R. Frand, E. Chitouras, and C. A. Kaiser A Link between Secretion and Pre-mRNA Processing Defects in Saccharomyces cerevisiae and the Identification of a Novel Splicing Gene, RSE1 Mol. Cell. Biol., December 1, 1998; 18(12): 7139 - 7146. [Abstract] [Full Text]
|
|
|
|
|
|
O. Gozani, J. Potashkin, and R. Reed A Potential Role for U2AF-SAP 155 Interactions in Recruiting U2 snRNP to the Branch Site Mol. Cell. Biol., August 1, 1998; 18(8): 4752 - 4760. [Abstract] [Full Text]
|
|
|
|
|
|
S. W. Ruby Dynamics of the U1 Small Nuclear Ribonucleoprotein during Yeast Spliceosome Assembly J. Biol. Chem., July 11, 1997; 272(28): 17333 - 17341. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. L. O'Day, G. Dalbadie-McFarland, and J. Abelson The Saccharomyces cerevisiae Prp5 Protein Has RNA-dependent ATPase Activity with Specificity for U2 Small Nuclear RNA J. Biol. Chem., December 27, 1996; 271(52): 33261 - 33267. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Awasthi, R. Palmer, M. Castro, C. D. Mobarak, and S. W. Ruby New Roles for the Snp1 and Exo84 Proteins in Yeast Pre-mRNA Splicing J. Biol. Chem., August 10, 2001; 276(33): 31004 - 31015. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |