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PMID: 7962083 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of nuclear polyadenylated RNA-binding proteins in Saccharomyces cerevisiae.

The Journal of cell biology ·Vol. 127 ·No. 5 ·1994-12-00 ·Pages 1173-84

Wilson SM, Datar KV, Paddy MR, Swedlow JR, Swanson MS

Abstract

To study the functions of heterogeneous nuclear ribonucleoproteins (hnRNPs), we have characterized nuclear polyadenylated RNA-binding (Nab) proteins from Saccharomyces cerevisiae. Nab1p, Nab2p, and Nab3p were isolated by a method which uses UV light to cross-link proteins directly bound to poly(A)+ RNA in vivo. We have previously characterized Nab2p, and demonstrated that it is structurally related to human hnRNPs. Here we report that Nab1p is identical to the Np13p/Nop3p protein recently implicated in both nucleocytoplasmic protein shuttling and pre-rRNA processing, and characterize a new nuclear polyadenylated RNA-binding protein, Nab3p. The intranuclear distributions of the Nab proteins were analyzed by three-dimensional immunofluorescence optical microscopy. All three Nab proteins are predominantly localized within the nucleoplasm in a pattern similar to the distribution of hnRNPs in human cells. The NAB3 gene is essential for cell viability and encodes an acidic ribonucleoprotein. Loss of Nab3p by growth of a GAL::nab3 mutant strain in glucose results in a decrease in the amount of mature ACT1, CYH2, and TPI1 mRNAs, a concomitant accumulation of unspliced ACT1 pre-mRNA, and an increase in the ratio of unspliced CYH2 pre-mRNA to mRNA. These results suggest that the Nab proteins may be required for packaging pre-mRNAs into ribonucleoprotein structures amenable to efficient nuclear RNA processing.

MeSH Terms
Amino Acid Sequence Base Sequence Cell Division Cell Nucleus/chemistry Cell Survival Fungal Proteins/chemistry,genetics,isolation & purification,metabolism Genes, Fungal Heterogeneous-Nuclear Ribonucleoproteins Molecular Sequence Data Nuclear Proteins/chemistry,genetics,isolation & purification,metabolism Nucleocytoplasmic Transport Proteins RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA, Fungal/metabolism RNA, Heterogeneous Nuclear/metabolism RNA, Messenger/metabolism RNA-Binding Proteins/chemistry,genetics,isolation & purification,metabolism Ribonucleoproteins/chemistry Saccharomyces cerevisiae/chemistry,genetics,growth & development Saccharomyces cerevisiae Proteins
Chemicals
Fungal Proteins Heterogeneous-Nuclear Ribonucleoproteins NAB2 protein, S cerevisiae NAB3 protein, S cerevisiae NPL3 protein, S cerevisiae Nuclear Proteins Nucleocytoplasmic Transport Proteins RNA Precursors RNA, Fungal RNA, Heterogeneous Nuclear RNA, Messenger RNA-Binding Proteins Ribonucleoproteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wilson S M
Department of Immunology and Medical Microbiology, University of Florida, College of Medicine, Gainesville 32610-0266.
Datar K V
Paddy M R
Swedlow J R
Swanson M S
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.127.5.1173
Published
1994-12-00
Pages
1173-84
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120247
Subset
IM
Grants
NIGMS NIH HHS · GM46272 · United States
Databases
GENBANK
U05314
External Links
PubMed source DOI: 10.1083/jcb.127.5.1173 PMC Full Text View journal details
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