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Nucleic Acids Research, 1980, Vol. 8, No. 14 3065-3072
© 1980

MOLECULAR BIOLOGY

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

Peter Westermann, Odd Nygard* and Heinz Bielka

Central Institute of Molecular Biology, Academy of Sciences of GDR 1115 Berlin-Buch, GDR, Sweden *Department of Cell Physiology, The Wenner-Gren Institute, University of Stockholm S-113 45 Stockholm, Sweden

Received June 13, 1980. Initiation factor eIF-2 from rat liver was reacted with the heterobifunctional 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.


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