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Journal of Bacteriology, May 2004, p. 3046-3055, Vol. 186, No. 10
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.10.3046-3055.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Temperature Sensitivity Caused by Mutant Release Factor 1 Is Suppressed by Mutations That Affect 16S rRNA Maturation

Magdalena Kaczanowska and Monica Rydén-Aulin*

Department of Genetics, Microbiology and Toxicology, University of Stockholm, S-106 91 Stockholm, Sweden

Received 10 December 2003/ Accepted 6 February 2004

To study the effect of slow termination on the protein synthesizing machinery, we isolated suppressors to a temperature-sensitive release factor 1 (RF1). Of 26 independent clones, five complementation groups have been identified, two of which are presented here. The first mutation disrupts a base pair in the transcription terminator stem for the rplM-rpsI operon, which encodes ribosomal proteins L13 and S9. We have found that this leads to readthrough of the terminator and that lower levels of transcript (compared to the results seen with the wild type) are found in the cell. This probably leads to decreased expression of the two proteins. The second mutation is a small deletion of the yrdC open reading frame start site, and it is not likely that the protein is expressed. Both mutant strains show an increased accumulation of 17S rRNA (immature 16S rRNA). Maturation of 16S rRNA is dependent on proper assembly of the ribosomal proteins, a process that is disturbed when proteins are missing. The function of the YrdC protein is not known, but it is able to bind to double-stranded RNA; therefore, we suggest that it is an assembly factor important for 30S subunit biogenesis. On the basis of our findings, we propose that lesser amounts of S9 or a lack of YrdC causes the maturation defect. We have shown that as a consequence of the maturation defect, fewer 70S ribosomes and polysomes are formed. This and other results suggest that it is the lowered concentration of functional ribosomes that suppresses the temperature sensitivity caused by the mutant RF1.

* Corresponding author. Mailing address: Department of Genetics, Microbiology and Toxicology, University of Stockholm, S-106 91 Stockholm, Sweden. Phone: 46 8 164155. Fax: 46 8 6129552. E-mail: monica{at}gmt.su.se.

Journal of Bacteriology, May 2004, p. 3046-3055, Vol. 186, No. 10
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.10.3046-3055.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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