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PMID: 12897147 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cotranscriptional recruitment of the U1 snRNP to intron-containing genes in yeast.

Molecular and cellular biology ·Vol. 23 ·No. 16 ·2003-08-00 ·Pages 5768-79

Kotovic KM, Lockshon D, Boric L, Neugebauer KM

Abstract

Evidence that pre-mRNA processing events are temporally and, in some cases, mechanistically coupled to transcription has led to the proposal that RNA polymerase II (Pol II) recruits pre-mRNA splicing factors to active genes. Here we address two key questions raised by this proposal: (i) whether the U1 snRNP, which binds to the 5' splice site of each intron, is recruited cotranscriptionally in vivo and, (ii) if so, where along the length of active genes the U1 snRNP is concentrated. Using chromatin immunoprecipitation (ChIP) in yeast, we show that elevated levels of the U1 snRNP were specifically detected in gene regions containing introns and downstream of introns but not along the length of intronless genes. In contrast to capping enzymes, which bind directly to Pol II, the U1 snRNP was poorly detected in promoter regions, except in genes harboring promoter-proximal introns. Detection of the U1 snRNP was dependent on RNA synthesis and was abolished by intron removal. Microarray analysis revealed that intron-containing genes were preferentially selected by ChIP with the U1 snRNP. Thus, U1 snRNP accumulation at genes correlated with the presence and position of introns, indicating that introns are necessary for cotranscriptional U1 snRNP recruitment and/or retention.

MeSH Terms
Chromatin/metabolism Gene Deletion Genome Genotype Introns Models, Genetic Oligonucleotide Array Sequence Analysis Open Reading Frames Precipitin Tests Promoter Regions, Genetic Ribonucleoprotein, U1 Small Nuclear/metabolism Ribonucleoproteins, Small Nuclear/metabolism Saccharomyces cerevisiae/metabolism Temperature Transcription, Genetic
Chemicals
Chromatin Ribonucleoprotein, U1 Small Nuclear Ribonucleoproteins, Small Nuclear
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kotovic Kimberly M
Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
Lockshon Daniel
Boric Lamia
Neugebauer Karla M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.16.5768-5779.2003
Published
2003-08-00
Pages
5768-79
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC166328
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.23.16.5768-5779.2003 PMC Full Text View journal details
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