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Journal of Bacteriology, March 2009, p. 1604-1609, Vol. 191, No. 5
0021-9193/09/$08.00+0     doi:10.1128/JB.01485-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Hypomodification of the Wobble Base in tRNAGlu, tRNALys, and tRNAGln Suppresses the Temperature-Sensitive Phenotype Caused by Mutant Release Factor 1{triangledown}

Georgina Isak and Monica Rydén-Aulin*

Department of Genetics, Microbiology, and Toxicology, Stockholm University, S-10691 Stockholm, Sweden

Received 22 October 2008/ Accepted 9 December 2008

In Escherichia coli, release factor 1 (RF1) is one of two RFs that mediate termination; it specifically recognizes UAA and UAG stop codons. A mutant allele, prfA1, coding for an RF1 that causes temperature-sensitive (Ts) growth at 42°C, was used to select for temperature-resistant (Ts+) suppressors. This study describes one such suppressor that is the result of an IS10 insertion into the cysB gene, giving a Cys– phenotype. CysB is a transcription factor regulating the cys regulon, mainly as an activator, which explains the Cys– phenotype. We have found that suppression is a consequence of the lost ability to donate sulfur to enzymes involved in the synthesis of thiolated nucleosides. From genetic analyses we conclude that it is the lack of the 5-methylaminomethyl-2-thiouridine (mnm5s2U) modification of the wobble base of tRNAGlu, tRNALys, and/or tRNAGln that causes the suppressor phenotype.

* Corresponding author. Mailing address: Department of Genetics, Microbiology, and Toxicology, Stockholm University, S-10691 Stockholm, Sweden. Phone: 46-8-164154. Fax: 46-8-164315. E-mail: monica{at}gmt.su.se

{triangledown} Published ahead of print on 19 December 2008.

Journal of Bacteriology, March 2009, p. 1604-1609, Vol. 191, No. 5
0021-9193/09/$08.00+0     doi:10.1128/JB.01485-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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