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PMID: 8413213 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

PUB1: a major yeast poly(A)+ RNA-binding protein.

Molecular and cellular biology ·Vol. 13 ·No. 10 ·1993-10-00 ·Pages 6114-23

Matunis MJ, Matunis EL, Dreyfuss G

Abstract

The expression of RNA polymerase II transcripts can be regulated at the posttranscriptional level by RNA-binding proteins. Although extensively characterized in metazoans, relatively few RNA-binding proteins have been characterized in the yeast Saccharomyces cerevisiae. Three major proteins are cross-linked by UV light to poly(A)+ RNA in living S. cerevisiae cells. These are the 72-kDa poly(A)-binding protein and proteins of 60 and 50 kDa (S.A. Adam, T.Y. Nakagawa, M.S. Swanson, T. Woodruff, and G. Dreyfuss, Mol. Cell. Biol. 6:2932-2943, 1986). Here, we describe the 60-kDa protein, one of the major poly(A)+ RNA-binding proteins in S. cerevisiae. This protein, PUB1 [for poly(U)-binding protein 1], was purified by affinity chromatography on immobilized poly(rU), and specific monoclonal antibodies to it were produced. UV cross-linking demonstrated that PUB1 is bound to poly(A)+ RNA (mRNA or pre-mRNA) in living cells, and it was detected primarily in the cytoplasm by indirect immunofluorescence. The gene for PUB1 was cloned and sequenced, and the sequence was found to predict a 51-kDa protein with three ribonucleoprotein consensus RNA-binding domains and three glutamine- and asparagine-rich auxiliary domains. This overall structure is remarkably similar to the structures of the Drosophila melanogaster elav gene product, the human neuronal antigen HuD, and the cytolytic lymphocyte protein TIA-1. Each of these proteins has an important role in development and differentiation, potentially by affecting RNA processing. PUB1 was found to be nonessential in S. cerevisiae by gene replacement; however, further genetic analysis should reveal important features of this class of RNA-binding proteins.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular DNA, Fungal Drosophila melanogaster Fluorescent Antibody Technique Fungal Proteins/genetics,isolation & purification,metabolism Humans Immunoblotting Molecular Sequence Data Poly(A)-Binding Proteins Precipitin Tests RNA, Fungal/metabolism RNA-Binding Proteins/genetics,isolation & purification,metabolism Saccharomyces cerevisiae/genetics,growth & development,metabolism Saccharomyces cerevisiae Proteins
Chemicals
DNA, Fungal Fungal Proteins PUB1 protein, S cerevisiae Poly(A)-Binding Proteins RNA, Fungal RNA-Binding Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Matunis M J
Howard Hughes Medical Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6148.
Matunis E L
Dreyfuss G
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.10.6114-6123.1993
Published
1993-10-00
Pages
6114-23
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364671
Subset
IM
Databases
GENBANK
L20767
External Links
PubMed source DOI: 10.1128/mcb.13.10.6114-6123.1993 PMC Full Text View journal details
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