Isolation and characterization of an estrogen-regulated ribosome-associated inactivator of tRNA aminoacylation in the uterus.
Estradiol (E2) induces an increase in the peptide elongation rate of isolated uterine ribosomes assayed in a cell-free protein synthesis system. An inhibitory factor, extracted from ribosomes of E2-deprived rats, was found to inhibit the peptide elongation reaction by acting on certain tRNAs to render them incapable of binding to aminoacyl-tRNA synthetases, thus reducing the availability of specific aminoacylated tRNAs required for the sequential translation of the codons in mRNA. The uterine ribosome-associated tRNA inactivator (RATI) has been partially purified and monoclonal antibodies (MABs) to RATI have been prepared. Specificity of the MABs for RATI was indicated by the inactivation of RATI in vitro by the anti-RATI MABs. RATI selectively inactivates deacylated, but not acylated, tRNAs and the inactivation does not appear to involve nuclease cleavage of the tRNA. Within 1 h after E2 treatment 50% of both RATI activity and immunoreactivity were lost from the uterine ribosome extracts, suggesting that E2 regulation of tRNA reutilization may occur through dissociation of RATI from the ribosomal site of tRNA deacylation or alteration in the structure of RATI resulting in inactivation both biologically and immunologically. We propose that RATI may function as an E2-regulatable 'switch' mechanism which inactivates, delays or defers the aminoacylation of certain tRNAs in the absence of E2 and which participates in the regulation of protein synthesis at the translational level by creating rate-limiting levels of certain tRNAs in the E2-deprived uterus.
Whelly
S M
SM
Department of Biochemistry and Molecular Biology, Texas Tech University Health Sciences Center, Lubbock.
Rasmussen
K R
KR
Skudlarek
J
J
Barker
K L
KL
D000215:Acylation; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology; D000918:Antibody Specificity; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D012343:RNA, Transfer / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051381:Rats; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism; D014599:Uterus / Q000737:chemistry* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D004958:Estradiol; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases