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PMID: 2304466 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A multicomponent complex is required for the AAUAAA-dependent cross-linking of a 64-kilodalton protein to polyadenylation substrates.

Molecular and cellular biology ·Vol. 10 ·No. 3 ·1990-03-00 ·Pages 1244-8

Wilusz J, Shenk T, Takagaki Y, Manley JL

Abstract

A 64-kilodalton (kDa) polypeptide that is cross-linked by UV light specifically to polyadenylation substrate RNAs containing a functional AAUAAA element has been identified previously. Fractionated HeLa nuclear components that can be combined to regenerate efficient and accurate polyadenylation in vitro have now been screened for the presence of the 64-kDa protein. None of the individual components contained an activity which could generate the 64-kDa species upon UV cross-linking in the presence of substrate RNA. It was necessary to mix two components, cleavage stimulation factor and specificity factor, to reconstitute 64-kDa protein-RNA cross-linking. The addition of cleavage factors to this mixture very efficiently reconstituted the AAUAAA-specific 64-kDa protein-RNA interaction. The 64-kDa protein, therefore, is present in highly purified, reconstituted polyadenylation reactions. However, it is necessary to form a multicomponent complex to efficiently cross-link the protein to a substrate RNA.

MeSH Terms
Base Sequence Cell-Free System HeLa Cells Humans In Vitro Techniques Molecular Weight Nuclear Proteins/physiology Poly A/metabolism RNA Processing, Post-Transcriptional Ribonucleoproteins/physiology,radiation effects,ultrastructure Ultraviolet Rays
Chemicals
Nuclear Proteins Ribonucleoproteins Poly A
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wilusz J
Howard Hughes Medical Institute, Department of Biology, Princeton University, New Jersey 08544.
Shenk T
Takagaki Y
Manley J L
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.3.1244-1248.1990
Published
1990-03-00
Pages
1244-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361011
Subset
IM
Grants
NCI NIH HHS · CA-38965 · United States
NIGMS NIH HHS · GM-28983 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.3.1244-1248.1990 PMC Full Text View journal details
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