Escherichia coli Ribosomes as a Model for Testing New Photoactivated tRNA Analogs Containing 6-Thioguanosine Residues

A comparative study of photoreactive tRNA derivatives containing a 6-thioguanosine or diazirine derivative of 5-methyleneaminouridine was performed using Escherichia coli ribosomes. The conditions of producing functional tRNAs containing photoreactive residues have been selected using T7 RNA polymer...

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Bibliographic Details
Published in:Molecular biology (New York) 2004-09, Vol.38 (5), p.799-805
Main Authors: Lavrik, IN, Sergiev, P V, Bogdanov, A A, Zimmermann, R A, Dontsova, O A
Format: Article
Language:English
Subjects:
Comparative studies
DNA-directed RNA polymerase
E coli
Escherichia coli
Hydrolysis
Microbiology
Nucleotide analogs
Reverse transcription
Ribonuclease H
Ribosomes
RNA polymerase
rRNA
Transcription
tRNA
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Summary:A comparative study of photoreactive tRNA derivatives containing a 6-thioguanosine or diazirine derivative of 5-methyleneaminouridine was performed using Escherichia coli ribosomes. The conditions of producing functional tRNAs containing photoreactive residues have been selected using T7 RNA polymerase transcription. Photochemical modification was conducted upon binding the corresponding tRNA super(Phe) transcripts carrying the photoreactive residues of nucleotide analogs to the P site of the ribosome. The results of modification were analyzed using oligonucleotide-directed hydrolysis of rRNA by RNase H, as well as reverse transcription. It is shown that 6-thioguanosine is a promising agent for studying RNA-protein complexes by photochemical crosslinking methods.
ISSN:0026-8933
1608-3245
DOI:10.1023/B:MBIL.0000043949.56555.03