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PMID: 2961980 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Sedimentation analysis of polyadenylation-specific complexes.

Molecular and cellular biology ·Vol. 8 ·No. 1 ·1988-01-00 ·Pages 226-33

Moore CL, Skolnik-David H, Sharp PA

Abstract

Precursor RNA containing the adenovirus L3 polyadenylation site is assembled into a 50S complex upon incubation with HeLa nuclear extract at 30 degrees C. The cofactor and sequence requirements for 50S complex formation are similar to those of the in vitro polyadenylation reaction. Assembly of this complex requires ATP but is not dependent upon synthesis of a poly(A) tract. In addition, a 50S complex does not form on substrate RNA in which the AAUAAA hexanucleotide upstream of the poly(A) site has been mutated to AAGAAA or on RNA in which sequences between +5 and +48 nucleotides downstream of the site have been removed. These mutations also prevent in vitro processing of substrate RNA. Kinetic studies suggest that the 50S complex is an intermediate in the polyadenylation reaction. It forms at an early stage in the reaction and at later times contains both poly(A)+ RNA as well as unreacted precursor. U-type small nuclear ribonucleoprotein particles are components of the 50S complex, as shown by immunoprecipitation with antiserum specific to the trimethyl cap of these small nuclear RNAs.

MeSH Terms
Adenoviruses, Human/genetics Cell-Free System Centrifugation HeLa Cells In Vitro Techniques Macromolecular Substances Poly A/metabolism RNA Processing, Post-Transcriptional RNA, Messenger/metabolism Ribonucleoproteins/isolation & purification,metabolism Ribonucleoproteins, Small Nuclear Structure-Activity Relationship
Chemicals
Macromolecular Substances RNA, Messenger Ribonucleoproteins Ribonucleoproteins, Small Nuclear Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Moore C L
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Skolnik-David H
Sharp P A
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34 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.8.1.226-233.1988
Published
1988-01-00
Pages
226-33
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC363106
Subset
IM
Grants
NIGMS NIH HHS · GM32467 · United States
NCI NIH HHS · P01-CA42063 · United States
NCI NIH HHS · P30-CA14051 · United States
External Links
PubMed source DOI: 10.1128/mcb.8.1.226-233.1988 PMC Full Text View journal details
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