Determinants of Nam8-dependent splicing of meiotic pre-mRNAs Journal Article

   


Authors: Qiu, Z. R.; Schwer, B.; Shuman, S.
Article Title: Determinants of Nam8-dependent splicing of meiotic pre-mRNAs
Abstract: Nam8, a component of yeast U1 snRNP, is optional for mitotic growth but required during meiosis, because Nam8 collaborates with Mer1 to promote splicing of essential meiotic mRNAs AMA1, MER2 and MER3. Here, we identify SPO22 and PCH2 as novel targets of Nam8-dependent meiotic splicing. Whereas SPO22 splicing is co-dependent on Mer1, PCH2 is not. The SPO22 intron has a non-consensus 5' splice site (5'SS) that dictates its Nam8/Mer1-dependence. SPO22 splicing relies on Mer1 recognition, via its KH domain, of an intronic enhancer 5'-AYACCCUY. Mutagenesis of KH and the enhancer highlights Arg214 and Gln243 and the CCC triplet as essential for Mer1 activity. The Nam8-dependent PCH2 pre-mRNA has a consensus 5'SS and lacks a Mer1 enhancer. For PCH2, a long 5' exon and a non-consensus intron branchpoint dictate Nam8-dependence. Our results implicate Nam8 in two distinct meiotic splicing regulons. Nam8 is composed of three RRM domains, flanked by N-terminal leader and C-terminal tail segments. The leader, tail and RRM1 are dispensable for splicing meiotic targets and unnecessary for vegetative Nam8 function in multiple synthetic lethal genetic backgrounds. Nam8 activity is enfeebled by alanine mutations in the putative RNA binding sites of the RRM2 and RRM3 domains. © 2010 The Author(s).
Keywords: controlled study; protein expression; unclassified drug; gene mutation; nonhuman; protein domain; protein function; meiosis; reverse transcription polymerase chain reaction; intron; saccharomyces cerevisiae; binding site; alanine; saccharomyces cerevisiae protein; rna structure; small nuclear ribonucleoprotein; glycine; arginine; rna binding; rna splicing; complementary dna; rna sequence; messenger rna precursor; open reading frame; consensus sequence; rna isolation; nam8 protein; protein mer1; protein pch2; protein spo22; sporogenesis
Journal Title: Nucleic Acids Research
Volume: 39
Issue: 8
ISSN: 0305-1048
Publisher: Oxford University Press  
Date Published: 2011-04-01
Start Page: 3427
End Page: 3445
Language: English
DOI: 10.1093/nar/gkq1328
PROVIDER: scopus
PMCID: PMC3082912
PUBMED: 21208980
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-79955595852&partnerID=40&md5=e9d0421302fd818b2d6e9512d6aeec95
Notes: --- - "Export Date: 23 June 2011" - "CODEN: NARHA" - "Source: Scopus"
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MSK Authors
  1. Stewart H Shuman
    546 Shuman
  2. Zhicheng Qiu
    6 Qiu
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