Antibody-Nucleic Acid Complexes. Inhibition of Translation of Silkmoth Chorion Messenger Ribonucleic Acid with Antibodies Specific for 7-Methylguanosine

Theodore W. Munns, Charles S. Morrow, James R. Hunsley, Robert J. Oberst, M. Kathryn Liszewski

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Antibodies specific for 7-methylguanosine (m7G) were evaluated for their ability to inhibit the translation of chorion mRNA in a wheat germ cell-free amino acid incorporating system. Results obtained with antibody concentrations of 0.5-1.5 μM revealed dose-dependent inhibition of [3H]-labeled amino acid incorporation into acid-insoluble radioactivity. Inhibition of translation was attributed to the interaction of anti-m7G antibodies with the 5' termini of chorion mRNAs on the basis that (a) anti-m7G antibodies coupled to Sepharose (anti-m7G-Sepharose) immunospecif-ically retained 5'-terminal cap structures of chorion mRNAs m7G (5')ppp(5')Nm (b) significant inhibition of translation required a 2-h preincubation of anti-m7G antibodies with i.e., mRNA, and (c) similar preincubation periods with anti-m7G antibodies in the presence of the competing nucleoside hapten (m7G) obviated the inhibitory effect of the antibody. The nature of the anti-m7G antibody-mRNA complex was examined by digesting chorion mRNA with nuclease P, before (predigested) and after (postdigested) immunospecific adsorption to anti-m7G-Sepharose adsorbent. Whereas predigested preparations yielded a single cap structure of the type m7G(5')ppp(5')N, the predominating cap in the postdigested sample was m7G(5')ppp(5')NpNpN. These latter data revealed that the nucleotide sequence adjacent to the cap was not significantly masked by the antibody and suggest the utility of anti-m7G antibody as a site-specific probe.

Original languageEnglish
Pages (from-to)3804-3810
Number of pages7
JournalBiochemistry
Volume18
Issue number17
DOIs
StatePublished - Jan 1 1979

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