UV cross-linking of polypeptides associated with 3'-terminal exons

Journal Article · Thu Nov 01 04:00:00 EST 1990 · Molecular and Cellular Biology; (USA)

DOI:https://doi.org/10.1128/MCB.10.11.5937· OSTI ID:6168327

Stolow, D T; Berget, S M ^[1]

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1. Baylor College of Medicine, Houston, TX (USA)

Association of nuclear proteins with chimeric vertebrate precursor RNAs containing both polyadenylation signals and an intron was examined by UV cross-linking. One major difference in cross-linking pattern was observed between this chimeric precursor RNA and precursors containing only polyadenylation or splicing signals. The heterogeneous nuclear ribonucleoprotein (hnRNP) polypeptide C cross-linked strongly to sequences downstream of the A addition site in polyadenylation precursor RNA containing only the polyadenylation signal from the simian virus 40 (SV40) late transcription unit. In contrast, the hnRNP C polypeptide cross-linked to chimeric RNA containing the same SV40 late poly(A) cassette very poorly, at a level less than 5% of that observed with the precursor RNA containing just the poly(A) site. Observation that cross-linking of the hnRNP C polypeptide to elements within the SV40 late poly(A) site was altered by the presence of an upstream intron suggests differences in the way nuclear factors associate with poly(A) sites in the presence and absence of an upstream intron. Cross-linking of C polypeptide to chimeric RNA increased with RNAs mutated for splicing or polyadenylation consensus sequences and under reaction conditions (high magnesium) that inhibited polyadenylation. Furthermore, cross-linking of hnRNP C polypeptide to precursors containing just the SV40 late poly(A) site was eliminated in the presence of competing poly(U); polyadenylation, however, was unaffected. Correlation of loss of activity with high levels of hnRNP C polypeptide cross-linking raises questions about the specificity of the interaction between the hnRNP C polypeptide and polyadenylation precursor RNAs in vitro.

• OSTI ID: 6168327
• Journal Information: Molecular and Cellular Biology; (USA), Journal Name: Molecular and Cellular Biology; (USA) Vol. 10:11; ISSN MCEBD; ISSN 0270-7306
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
CHEMICAL REACTIONS
CROSS-LINKING
ELECTROMAGNETIC RADIATION
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PARASITES
PEPTIDES
POLYMERIZATION
POLYPEPTIDES
PRECURSOR
PROTEINS
RADIATIONS
RNA
SIMIAN VIRUS
TRANSCRIPTION
ULTRAVIOLET RADIATION
VIRUSES