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:
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