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PMID: 9649443 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Depletion of yeast RNase III blocks correct U2 3' end formation and results in polyadenylated but functional U2 snRNA.

The EMBO journal ·Vol. 17 ·No. 13 ·1998-07-01 ·Pages 3738-46

Abou Elela S, Ares M

Abstract

Yeast U2 snRNA is transcribed by RNA polymerase II to generate a single non-polyadenylated transcript. A temperature-sensitive yeast strain carrying a disruption in RNT1, the gene encoding a homolog of RNase III, produces 3'-extended U2 that is polyadenylated. The U2 3'-flanking region contains a putative stem-loop that is recognized and cleaved at two sites by recombinant GST-Rnt1 protein in vitro. Removal of sequences comprising the stem-loop structure blocks cleavage in vitro and mimics the effects of Rnt1 depletion in vivo. Strains carrying a U2 gene lacking the Rnt1 cleavage site produce only polyadenylated U2 snRNA, and yet are not impaired in growth or splicing. The results suggest that eukaryotic RNase III may be a general factor in snRNA processing, and demonstrate that polyadenylation is not incompatible with snRNA function in yeast.

MeSH Terms
Base Sequence Cell Division Endoribonucleases/genetics,metabolism Glutathione Transferase/genetics,metabolism Molecular Sequence Data Mutagenesis Nucleic Acid Conformation Poly A RNA Splicing RNA, Fungal/chemistry Recombinant Fusion Proteins/genetics,metabolism Ribonuclease III Ribonucleoprotein, U2 Small Nuclear/metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Structure-Activity Relationship
Chemicals
RNA, Fungal Recombinant Fusion Proteins Ribonucleoprotein, U2 Small Nuclear Saccharomyces cerevisiae Proteins Poly A Glutathione Transferase Endoribonucleases RNT1 protein, S cerevisiae Ribonuclease III
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Abou Elela S
Center for the Molecular Biology of RNA, Biology Department, Sinsheimer Laboratories, University of California at Santa Cruz, Santa Cruz, CA 95064, USA. [email protected]
Ares M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.13.3738
Email
[email protected]
Published
1998-07-01
Pages
3738-46
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170709
Subset
IM
Grants
NIGMS NIH HHS · GM 55557 · United States
External Links
PubMed source DOI: 10.1093/emboj/17.13.3738 PMC Full Text View journal details
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