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PMID: 2408761 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Accurate cleavage and polyadenylation of exogenous RNA substrate.

Cell ·Vol. 41 ·No. 3 ·1985-07-00 ·Pages 845-55

Moore CL, Sharp PA

Abstract

Purified precursor RNA containing the L3 polyadenylation site of late adenovirus 2 mRNA is accurately cleaved and polyadenylated when incubated with nuclear extract from HeLa cells. The reaction is very efficient; 75% of the precursor is correctly processed. Cleavage is rapidly followed by polymerization of an initial poly(A) tract of approximately 130 nucleotides. Additional adenosine residues are added during further incubation. In the presence of the ATP analog alpha-beta-methylene-adenosine 5' triphosphate, the precursor RNA is cleaved but not polyadenylated, suggesting that processing is not coupled to the synthesis of the initial poly(A) tract. In the absence of free Mg2+, a small RNA of approximately 46 nucleotides is stabilized against degradation. Fingerprint analysis suggests this RNA is produced by endonucleolytic cleavage at the L3 site. Like the in vitro splicing reaction, the in vitro polyadenylation reaction is inhibited by adding antiserum against the small nuclear ribonucleoprotein particle containing U1 RNA.

MeSH Terms

Adenosine Triphosphate/pharmacology Adenoviruses, Human Base Sequence HeLa Cells Humans Kinetics Magnesium/pharmacology Nucleic Acid Precursors/metabolism Poly A/metabolism RNA/metabolism RNA Caps/physiology RNA Precursors RNA Processing, Post-Transcriptional RNA, Messenger RNA, Viral/metabolism Ribonucleoproteins/physiology Ribonucleoproteins, Small Nuclear

Chemicals

Nucleic Acid Precursors RNA Caps RNA Precursors RNA, Messenger RNA, Viral Ribonucleoproteins Ribonucleoproteins, Small Nuclear Poly A RNA Adenosine Triphosphate Magnesium

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Moore C L

Sharp P A

Article Info

Journal

Cell

Abbr.

Cell

ISSN

0092-8674

DOI

10.1016/s0092-8674(85)80065-9

Published

1985-07-00

Pages

845-55

Language

English

Region

United States

NLM ID

0413066

Subset

Grants

NIGMS NIH HHS · 5 F32-GM08770 · United States

NCI NIH HHS · P01-CA26717 · United States

NIGMS NIH HHS · R01-GM32467 · United States

Databases

GENBANK

J01917, J01918, J01919, J01920, J01921, J01922, J01923, J01924, J01925, J01926, J01927, J01928, J01929, J01930, J01931, J01932, J01933, J01934, J01935, J01936, J01937, J01938, J01939, J01940, J01941, J01942, J01943, J01944, J01945, J01946

External Links

PubMed source DOI: 10.1016/s0092-8674(85)80065-9 View journal details

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Accurate cleavage and polyadenylation of exogenous RNA substrate.

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