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A functional site of the GTPase‐associated center within 28S ribosomal RNA probed with an anti‐RNA autoantibody.
Author(s) -
Uchiumi T.,
Kominami R.
Publication year - 1994
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1994.tb06641.x
Subject(s) - biology , rna , 28s ribosomal rna , ribosomal rna , microbiology and biotechnology , ribosome , genetics , virology , gene
An anti‐RNA autoantibody (anti‐28S) was employed to identify structural and functional elements characteristic of a domain termed the ‘GTPase center’ in eukaryotic 28S ribosomal RNA. This antibody, an inhibitor of ribosome‐associated GTP hydrolysis, has a unique property: it binds to the RNA domain of eukaryotes but not to that of prokaryotes. The antibody binding occurred in the presence of Mg2+ and protected from chemical modification three conserved bases (U1958, G1960 and A1990) and the base G1959 which is replaced by A in prokaryotic 23S rRNA (A1067 in Escherichia coli). In vitro substitution of G1959 to A drastically weakened the antibody binding, and the reciprocal substitution, A1067‐‐>G of the E.coli domain conferred the binding ability. This suggests that the G base determines the specificity of antibody binding. The G1959 was also protected by the association of ribosomes with elongation factor EF‐2. The result, together with protection of E.coli base A1067 by EFG [D.Moazed, I.M.Robertson and H.F.Noller (1988) Nature, 334, 362‐364], suggests that the position of G1959 in eukaryotes and A1067 in prokaryotes constitutes at least part of the factor binding site irrespective of the base replacement during evolution.