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Molecular and Cellular Biology, October 2004, p. 9186-9197, Vol. 24, No. 20
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.20.9186-9197.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Ribosome-Bound Chaperones RAC and Ssb1/2p Are Required for Accurate Translation in Saccharomyces cerevisiae

Magdalena Rakwalska and Sabine Rospert*

Institut für Biochemie und Molekularbiologie, Universität Freiburg, Freiburg, Germany

Received 26 April 2004/ Returned for modification 19 May 2004/ Accepted 22 July 2004

The chaperone homologs RAC (ribosome-associated complex) and Ssb1/2p are anchored to ribosomes; Ssb1/2p directly interacts with nascent polypeptides. The absence of RAC or Ssb1/2p results in a similar set of phenotypes, including hypersensitivity against the aminoglycoside paromomycin, which binds to the small ribosomal subunit and compromises the fidelity of translation. In order to understand this phenomenon we measured the frequency of translation termination and misincorporation in vivo and in vitro with a novel reporter system. Translational fidelity was impaired in the absence of functional RAC or Ssb1/2p, and the effect was further enhanced by paromomycin. The mutant strains suffered primarily from a defect in translation termination, while misincorporation was compromised to a lesser extent. Consistently, a low level of soluble translation termination factor Sup35p enhanced growth defects in the mutant strains. Based on the combined data we conclude that RAC and Ssb1/2p are crucial in maintaining translational fidelity beyond their postulated role as chaperones for nascent polypeptides.

* Corresponding author. Mailing address: Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herder-Strasse 7, D-79104 Freiburg, Germany. Phone: 49-761-2035259. Fax: 49-761-2035257. E-mail: sabine.rospert{at}biochemie.uni-freiburg.de.

Molecular and Cellular Biology, October 2004, p. 9186-9197, Vol. 24, No. 20
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.20.9186-9197.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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