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PMID: 15929942 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

mRNA with a <20-nt poly(A) tail imparted by the poly(A)-limiting element is translated as efficiently in vivo as long poly(A) mRNA.

RNA (New York, N.Y.) ·Vol. 11 ·No. 7 ·2005-07-00 ·Pages 1131-40

Peng J, Schoenberg DR

Abstract

The poly(A)-limiting element (PLE) is a conserved sequence that restricts the length of the poly(A) tail to <20 nt. This study compared the translation of PLE-containing short poly(A) mRNA expressed in cells with translation in vitro of mRNAs with varying length poly(A) tails. In transfected cells, PLE-containing mRNA had a <20-nt poly(A) and accumulated to a level 20% higher than a matching control without a PLE. It was translated as well as the matching control mRNA with long poly(A) and showed equivalent binding to polysomes. Translation in a HeLa cell cytoplasmic extract was used to examine the impact of the PLE in the context of varying length poly(A) tails. Here the overall translation of +PLE mRNA was less than control mRNA with the same length poly(A), and the PLE did not overcome the effect of a short poly(A) tail. Because poly(A)-binding protein (PABP) is a dominant effector of poly(A)-dependent translation we reasoned excess PABP in our extract might overwhelm PLE regulation of translation. This was confirmed by experiments where PABP was inactivated with poly(rA) or Paip2, and the effect of both treatments was reversed by addition of recombinant PABP. These data indicate that the PLE functionally substitutes for bound PABP to stimulate translation of short poly(A) mRNA.

MeSH Terms
Animals Base Sequence COS Cells Cell Culture Techniques Chlorocebus aethiops Conserved Sequence HeLa Cells Humans Molecular Sequence Data Poly A/chemistry,genetics,metabolism Polyribosomes/metabolism Protein Biosynthesis RNA, Messenger/biosynthesis,chemistry,genetics RNA-Binding Proteins/biosynthesis,genetics Recombinant Proteins/biosynthesis
Chemicals
RNA, Messenger RNA-Binding Proteins Recombinant Proteins Poly A
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Peng Jing
Department of Molecular and Cellular Biochemistry, The Ohio State University, 1645 Neil Ave., Columbus, OH 43210-1218, USA.
Schoenberg Daniel R
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1261/rna.2470905
Published
2005-07-00
Epub
2005-00-31
Pages
1131-40
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1237109
Subset
IM
Grants
NIGMS NIH HHS · R01 GM038277 · United States
NIGMS NIH HHS · R01 GM38277 · United States
NIGMS NIH HHS · R01 GM055407 · United States
NIGMS NIH HHS · R01 GM55407 · United States
NCI NIH HHS · P30 CA16058 · United States
NCI NIH HHS · P30 CA016058 · United States
External Links
PubMed source DOI: 10.1261/rna.2470905 PMC Full Text View journal details
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