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PMID: 1701018 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A uridylate tract mediates efficient heterogeneous nuclear ribonucleoprotein C protein-RNA cross-linking and functionally substitutes for the downstream element of the polyadenylation signal.

Molecular and cellular biology ·Vol. 10 ·No. 12 ·1990-12-00 ·Pages 6397-407

Wilusz J, Shenk T

Abstract

Every RNA added to an in vitro polyadenylation extract became stably associated with both the heterogeneous nuclear ribonucleoprotein (hnRNP) A and C proteins, as assayed by immunoprecipitation analysis using specific monoclonal antibodies. UV-cross-linking analysis, however, which assays the specific spatial relationship of certain amino acids and RNA bases, indicated that the hnRNP C proteins, but not the A proteins, were associated with downstream sequences of the simian virus 40 late polyadenylation signal in a sequence-mediated manner. A tract of five consecutive uridylate residues was required for this interaction. The insertion of a five-base U tract into a pGEM4 polylinker-derived transcript was sufficient to direct sequence-specific cross-linking of the C proteins to RNA. Finally, the five-base uridylate tract restored efficient in vitro processing to several independent poly(A) signals in which it substituted for downstream element sequences. The role of the downstream element in polyadenylation efficiency, therefore, may be mediated by sequence-directed alignment or phasing of an hnRNP complex.

MeSH Terms
Base Sequence Cell Nucleus/metabolism,radiation effects HeLa Cells/metabolism,radiation effects Heterogeneous-Nuclear Ribonucleoprotein Group C Heterogeneous-Nuclear Ribonucleoproteins Humans Macromolecular Substances Molecular Sequence Data Oligonucleotide Probes Plasmids Poly A/genetics Protein Binding RNA/genetics RNA, Heterogeneous Nuclear/metabolism RNA, Messenger Restriction Mapping Ribonucleoproteins/metabolism Transcription, Genetic Ultraviolet Rays Uracil Uridine Triphosphate/metabolism
Chemicals
Heterogeneous-Nuclear Ribonucleoprotein Group C Heterogeneous-Nuclear Ribonucleoproteins Macromolecular Substances Oligonucleotide Probes RNA, Heterogeneous Nuclear RNA, Messenger Ribonucleoproteins Poly A Uracil RNA Uridine Triphosphate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wilusz J
Department of Microbiology and Molecular Genetics, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark 07103.
Shenk T
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.12.6397-6407.1990
Published
1990-12-00
Pages
6397-407
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362916
Subset
IM
Grants
NCRR NIH HHS · 2 S07 RR05393 · United States
NCI NIH HHS · CA-38965 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.12.6397-6407.1990 PMC Full Text View journal details
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