Suppression of UGA codon by a tryptophan tRNA

Teh sheng Chan, Robert E. Webster, Norton D. Zinder

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Tryptophan tRNA isolated from a UGA suppressor (SuUGA +) strain of Escherichia coli, CAJ64, has been partially purified on a benzoylated DEAE-cellulose column. When added to an in vitro system this tRNA was able to suppress a UGA mutation in the phage-specific polymerase of a mutant of bacteriophage f2 (op 9). Tryptophan tRNA obtained from an Su- strain did not cause suppression, nor did tRNA from the SuUGA + strain which had undergone a similar purification process but was enriched for tyrosine or phenylalanine acceptor activity. The suppressing tRNA was further fractionated and the suppressor activity correlated with tryptophan acceptor activity. We conclude that the suppressor tRNA species in E. coli CAJ64 is a tryptophan tRNA. The UGA mutant is known to be leaky. In order to facilitate the suppressor assay, use was made of an extract prepared from an Su- strr strain in which residual synthesis of polymerase stimulated by op 9 RNA was markedly reduced. We have demonstrated by mixing experiments that it is the ribosome fraction in the strr extract which is responsible for the observed streptomycin effect.

Original languageEnglish (US)
Pages (from-to)101-116
Number of pages16
JournalJournal of Molecular Biology
Volume56
Issue number1
DOIs
StatePublished - Feb 28 1971
Externally publishedYes

Fingerprint

Terminator Codon
Transfer RNA
Tryptophan
Bacteriophages
Escherichia coli
Streptomycin
Phenylalanine
Ribosomes
Tyrosine
RNA
Mutation

ASJC Scopus subject areas

  • Virology

Cite this

Suppression of UGA codon by a tryptophan tRNA. / Chan, Teh sheng; Webster, Robert E.; Zinder, Norton D.

In: Journal of Molecular Biology, Vol. 56, No. 1, 28.02.1971, p. 101-116.

Research output: Contribution to journalArticle

Chan, Teh sheng ; Webster, Robert E. ; Zinder, Norton D. / Suppression of UGA codon by a tryptophan tRNA. In: Journal of Molecular Biology. 1971 ; Vol. 56, No. 1. pp. 101-116.
@article{08f16e55b11f46ba9c76e6e70f4bf19b,
title = "Suppression of UGA codon by a tryptophan tRNA",
abstract = "Tryptophan tRNA isolated from a UGA suppressor (SuUGA +) strain of Escherichia coli, CAJ64, has been partially purified on a benzoylated DEAE-cellulose column. When added to an in vitro system this tRNA was able to suppress a UGA mutation in the phage-specific polymerase of a mutant of bacteriophage f2 (op 9). Tryptophan tRNA obtained from an Su- strain did not cause suppression, nor did tRNA from the SuUGA + strain which had undergone a similar purification process but was enriched for tyrosine or phenylalanine acceptor activity. The suppressing tRNA was further fractionated and the suppressor activity correlated with tryptophan acceptor activity. We conclude that the suppressor tRNA species in E. coli CAJ64 is a tryptophan tRNA. The UGA mutant is known to be leaky. In order to facilitate the suppressor assay, use was made of an extract prepared from an Su- strr strain in which residual synthesis of polymerase stimulated by op 9 RNA was markedly reduced. We have demonstrated by mixing experiments that it is the ribosome fraction in the strr extract which is responsible for the observed streptomycin effect.",
author = "Chan, {Teh sheng} and Webster, {Robert E.} and Zinder, {Norton D.}",
year = "1971",
month = "2",
day = "28",
doi = "10.1016/0022-2836(71)90087-8",
language = "English (US)",
volume = "56",
pages = "101--116",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Suppression of UGA codon by a tryptophan tRNA

AU - Chan, Teh sheng

AU - Webster, Robert E.

AU - Zinder, Norton D.

PY - 1971/2/28

Y1 - 1971/2/28

N2 - Tryptophan tRNA isolated from a UGA suppressor (SuUGA +) strain of Escherichia coli, CAJ64, has been partially purified on a benzoylated DEAE-cellulose column. When added to an in vitro system this tRNA was able to suppress a UGA mutation in the phage-specific polymerase of a mutant of bacteriophage f2 (op 9). Tryptophan tRNA obtained from an Su- strain did not cause suppression, nor did tRNA from the SuUGA + strain which had undergone a similar purification process but was enriched for tyrosine or phenylalanine acceptor activity. The suppressing tRNA was further fractionated and the suppressor activity correlated with tryptophan acceptor activity. We conclude that the suppressor tRNA species in E. coli CAJ64 is a tryptophan tRNA. The UGA mutant is known to be leaky. In order to facilitate the suppressor assay, use was made of an extract prepared from an Su- strr strain in which residual synthesis of polymerase stimulated by op 9 RNA was markedly reduced. We have demonstrated by mixing experiments that it is the ribosome fraction in the strr extract which is responsible for the observed streptomycin effect.

AB - Tryptophan tRNA isolated from a UGA suppressor (SuUGA +) strain of Escherichia coli, CAJ64, has been partially purified on a benzoylated DEAE-cellulose column. When added to an in vitro system this tRNA was able to suppress a UGA mutation in the phage-specific polymerase of a mutant of bacteriophage f2 (op 9). Tryptophan tRNA obtained from an Su- strain did not cause suppression, nor did tRNA from the SuUGA + strain which had undergone a similar purification process but was enriched for tyrosine or phenylalanine acceptor activity. The suppressing tRNA was further fractionated and the suppressor activity correlated with tryptophan acceptor activity. We conclude that the suppressor tRNA species in E. coli CAJ64 is a tryptophan tRNA. The UGA mutant is known to be leaky. In order to facilitate the suppressor assay, use was made of an extract prepared from an Su- strr strain in which residual synthesis of polymerase stimulated by op 9 RNA was markedly reduced. We have demonstrated by mixing experiments that it is the ribosome fraction in the strr extract which is responsible for the observed streptomycin effect.

UR - http://www.scopus.com/inward/record.url?scp=0015242936&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0015242936&partnerID=8YFLogxK

U2 - 10.1016/0022-2836(71)90087-8

DO - 10.1016/0022-2836(71)90087-8

M3 - Article

C2 - 4929882

AN - SCOPUS:0015242936

VL - 56

SP - 101

EP - 116

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 1

ER -