Poly(4‐thiouridylic Acid) as Messenger RNA and Its Application for Photoaffinity Labelling of the Ribosomal mRNA Binding Site
- 1 April 1977
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 74 (3), 447-456
- https://doi.org/10.1111/j.1432-1033.1977.tb11411.x
Abstract
Poly(4-thiouridylic acid) [poly(s4U)] synthesized by polymerization of 4-thiouridine 5''-diphosphate with Escherichia coli polynucleotide phosphorylase (EC 2.7.7.8) acts as mRNA in vitro in a protein-synthesizing system from E. coli. It stimulates binding of Phe-tRNA to ribosomes in the presence of EF[elongation factor]-Tu .cntdot. Ts at 5 mM Mg2+ concentration and nonenzymatically at 20 mM Mg2+ concentration. It codes for the synthesis of polyphenylalanine. Poly(s4U) competes with poly(U) for binding to E. coli ribosomes. Light of 330 nm photoactivates poly(s4U), making it a useful photoaffinity label for the ribosomal mRNA binding site. Upon irradiation of 70-S ribosomal complexes, photoreaction occurs with ribosomal proteins and 16-S RNA. Ribosomes preincubated with R17 RNA are protected against the photoaffinity reaction. The labeling of 16-S RNA can be reduced by treatment of ribosomes with colicin E3.This publication has 45 references indexed in Scilit:
- The codon binding site of the Escherichia coli ribosome as studied with a chemically reactive A-U-G analogJournal of Molecular Biology, 1975
- 30S Ribosomal proteins associated with the 3′‐terminus of 16S RNAFEBS Letters, 1975
- Identification of protein S 1 at the messenger RNA binding site of the Escherichia coli ribosomeBiochemical and Biophysical Research Communications, 1974
- A protein-nucleic acid crosslink in 30s ribosomesBiochemical and Biophysical Research Communications, 1973
- Chemical modification of the transfer RNA and polyuridylic acid binding sites of Escherichia coli 30 s ribosomal subunitsJournal of Molecular Biology, 1971
- Reaction of N-ethyl maleimide with the ribosomes of Escherichia coliJournal of Molecular Biology, 1971
- Photochemically induced transition of 4-thiouridine to uridine or uridine and cytidine in E. coli transfer ribonucleic acidBiochemical and Biophysical Research Communications, 1969
- Polypetide chain termination: An active processJournal of Molecular Biology, 1968
- The possibility of aggregation of ribosomal rna during hot phenol-SDS deproteinizationBiochemical and Biophysical Research Communications, 1967
- Studies on the mechanism of messenger ribonucleic acid attachment to ribosomesJournal of Molecular Biology, 1966