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PMID: 2565532 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro.

Molecular and cellular biology ·Vol. 9 ·No. 2 ·1989-02-00 ·Pages 659-70

Bernstein P, Peltz SW, Ross J

Abstract

Using an in vitro mRNA decay system, we investigated how poly(A) and its associated poly(A)-binding protein (PABP) affect mRNA stability. Cell extracts used in the decay reactions were depleted of functional PABP either by adding excess poly(A) competitor or by passing the extracts over a poly(A)-Sepharose column. Polyadenylated mRNAs for beta-globin, chloramphenicol acetyltransferase, and simian virus 40 virion proteins were degraded 3 to 10 times faster in reactions lacking PABP than in those containing excess PABP. The addition of purified Saccharomyces cerevisiae or human cytoplasmic PABP to PABP-depleted reactions stabilized the polyadenylated mRNAs. In contrast, the decay rates of nonpolyadenylated mRNAs were unaffected by PABP, indicating that both the poly(A) and its binding protein were required for maintaining mRNA stability. A nonspecific single-stranded binding protein from Escherichia coli did not restore stability to polyadenylated mRNA, and the stabilizing effect of PABP was inhibited by anti-PABP antibody. The poly(A) tract was the first mRNA segment to be degraded in PABP-depleted reactions, confirming that the poly(A)-PABP complex was protecting the 3' region from nucleolytic attack. These results indicate that an important function of poly(A), in conjunction with its binding protein, is to protect polyadenylated mRNAs from indiscriminate destruction by cellular nucleases. A model is proposed to explain how the stability of an mRNA could be affected by the stability of its poly(A)-PABP complex.

MeSH Terms
Binding Sites Binding, Competitive Carrier Proteins/metabolism Humans In Vitro Techniques Models, Biological Poly A/metabolism Poly(A)-Binding Proteins RNA, Fungal/metabolism RNA, Messenger/metabolism Saccharomyces cerevisiae/metabolism
Chemicals
Carrier Proteins Poly(A)-Binding Proteins RNA, Fungal RNA, Messenger Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bernstein P
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Peltz S W
Ross J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.2.659-670.1989
Published
1989-02-00
Pages
659-70
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362643
Subset
IM
Grants
NCI NIH HHS · CA07175 · United States
NCI NIH HHS · CA09135 · United States
NCI NIH HHS · CA23076 · United States
External Links
PubMed source DOI: 10.1128/mcb.9.2.659-670.1989 PMC Full Text View journal details
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