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PMID: 7443514 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The alpha and gamma subunits of initiation factor eIF-2 can be cross-linked to 18S ribosomal RNA within the quaternary initiation complex, eIF-2.Met-tRNAf.GDPCP.small ribosomal subunit.

Nucleic acids research ·Vol. 8 ·No. 14 ·1980-07-25 ·Pages 3065-71

Westermann P, Nygård O, Bielka H

Abstract

Initiation factor eIF-2 from rat liver was reacted with the hetero-bifunctional cross-linking reagents ABAI or APTPI without diminishing its ability to form the quaternary initiation complex with Met-tRNAf, GDPCP and the small ribosomal subunit. Upon irradiation with UV light, subunits alpha and gamma of eIF-2 became covalently linked to 18S ribosomal RNA. The subunits were identified electrophoretically after isolation of the covalent protein-rRNA complexes and subsequent degradation of the rRNA by nuclease and alkali treatments. The close proximity of the two factor subunits to sequences of ribosomal RNA within the quaternary complex could be confirmed in a second set of experiments using unmodified, 125I-labeled factor and diepoxybutane as cross-linking reagent.

MeSH Terms
Animals Chemical Phenomena Chemistry Cross-Linking Reagents Eukaryotic Initiation Factor-2 Guanosine Triphosphate/analogs & derivatives,metabolism,radiation effects Imidoesters Methionine Peptide Chain Initiation, Translational Proteins/metabolism,radiation effects RNA, Ribosomal/metabolism RNA, Transfer/metabolism,radiation effects RNA, Transfer, Amino Acyl Rats Ribosomes/metabolism Ultraviolet Rays
Chemicals
Cross-Linking Reagents Eukaryotic Initiation Factor-2 Imidoesters Proteins RNA, Ribosomal RNA, Transfer, Amino Acyl eIF-2 Met-tRNA(f)(met) GDPCP methyl-4-azidobenzoylaminoacetimidate methyl-5-(4-azidophenyl)-4,5-dithiapentanimidate Guanosine Triphosphate RNA, Transfer Methionine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Westermann P
Nygård O
Bielka H
References (14)
14 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/8.14.3065
Published
1980-07-25
Pages
3065-71
Language
English
Region
England
NLM ID
0411011
PMCID
PMC324355
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/8.14.3065 PMC Full Text View journal details
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