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PMID: 8622683 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Invariant U2 RNA sequences bordering the branchpoint recognition region are essential for interaction with yeast SF3a and SF3b subunits.

Molecular and cellular biology ·Vol. 16 ·No. 3 ·1996-03-00 ·Pages 818-28

Yan D, Ares M

Abstract

U2 small nuclear RNA (snRNA) contains a sequence (GUAGUA) that pairs with the intron branchpoint during splicing. This sequence is contained within a longer invariant sequence of unknown secondary structure and function that extends between U2 and I and stem IIa. A part of this region has been proposed to pair with U6 in a structure called helix III. We made mutations to test the function of these nucleotides in yeast U2 snRNA. Most single base changes cause no obvious growth defects; however, several single and double mutations are lethal or conditional lethal and cause a block before the first step of splicing. We used U6 compensatory mutations to assess the contribution of helix III and found that if it forms, helix III is dispensable for splicing in Saccharomyces cerevisiae. On the other hand, mutations in known protein components of the splicing apparatus suppress or enhance the phenotypes of mutations within the invariant sequence that connect the branchpoint recognition sequence to stem IIa. Lethal mutations in the region are suppressed by Cus1-54p, a mutant yeast splicing factor homologous to a mammalian SF3b subunit. Synthetic lethal interactions show that this region collaborates with the DEAD-box protein Prp5p and the yeast SF3a subunits Prp9p, Prp11p, and Prp21p. Together, the data show that the highly conserved RNA element downstream of the branchpoint recognition sequence of U2 snRNA in yeast cells functions primarily with the proteins that make up SF3 rather than with U6 snRNA.

MeSH Terms
Base Sequence Binding Sites Conserved Sequence Fungal Proteins/genetics,metabolism Molecular Sequence Data Mutation RNA Splicing RNA, Small Nuclear/genetics,metabolism RNA-Binding Proteins Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins
Chemicals
CUS1 protein, S cerevisiae Fungal Proteins RNA, Small Nuclear RNA-Binding Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Yan D
Biology Department, Sinsheimer Laboratories, University of California, Santa Cruz 95064, USA.
Ares M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.3.818
Published
1996-03-00
Pages
818-28
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231062
Subset
IM
Grants
NIGMS NIH HHS · GM40478 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1128/MCB.16.3.818 PMC Full Text View journal details
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