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PMID: 9570316 Published · ppublish English Journal Article

Genetic interaction between U6 snRNA and the first intron nucleotide in Saccharomyces cerevisiae.

RNA (New York, N.Y.) ·Vol. 4 ·No. 2 ·1998-02-00 ·Pages 167-80

Luukkonen BG, Séraphin B

Abstract

Nuclear pre-mRNA splicing necessitates specific recognition of the pre-mRNA splice sites. It is known that 5' splice site selection requires base pairing of U6 snRNA with intron positions 4-6. However, no factor recognizing the highly conserved 5' splice site GU has yet been identified. We have tested if the known U6 snRNA-pre-mRNA interaction could be extended to include the first intron nucleotides and the conserved 50GAG52 sequence of U6 snRNA. We observe that some combinations of 5' splice site and U6 snRNA mutations produce a specific synthetic block to the first splicing step. In addition, the U6-G52U allele can switch between two competing 5' splice sites harboring different nucleotides following the cleavage site. These results indicate that U6 snRNA position 52 interacts with the first nucleotide of the intron before 5' splice site cleavage. Some combinations of U6 snRNA and pre-mRNA mutations also blocked the second splicing step, suggesting a role for the corresponding nucleotides in a proofreading step before exon ligation. From studies in diverse organisms, various functions have been ascribed to the conserved U6 snRNA 47ACAGAG52 sequence. Our results suggest that these discrepancies might reflect variations between different experimental systems and point to an important conserved role of this sequence in the splicing reaction.

MeSH Terms
Base Sequence Binding Sites/genetics Conserved Sequence Introns Mutation RNA Precursors/genetics,metabolism RNA Splicing/genetics RNA, Fungal/genetics,metabolism RNA, Small Nuclear/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism
Chemicals
RNA Precursors RNA, Fungal RNA, Small Nuclear
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Luukkonen B G
European Molecular Biology Laboratory, Heidelberg, Germany.
Séraphin B
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
Published
1998-02-00
Pages
167-80
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1369605
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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