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MBC in Press, published online ahead of print December 28, 2005
Mol. Biol. Cell 10.1091/mbc.E05-07-0632

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Submitted on July 14, 2005
Revised on November 16, 2005
Accepted on December 20, 2005

Rck2 Is Required for Reprogramming of Ribosomes during Oxidative Stress

Swarna Swaminathan,*{dagger} Tomas Masek,*{ddagger} Claes Molin,* Martin Pospisek,{ddagger} and Per Sunnerhagen*

*Department of Cell and Molecular Biology, Lundberg Laboratory, Göteborg University, S-405 30 Göteborg, Sweden; {ddagger}Department of Genetics and Microbiology, Charles University, 116 34 Prague 1, Czech Republic

Monitoring Editor: Thomas Fox

Rck2 is a MAP kinase-regulated protein kinase in yeast implicated in translational regulation. rck2{Delta} mutants are mildly sensitive to oxidative stress, a condition which causes dissociation of actively translating ribosomes (polysomes). In rck2{Delta} cells, polysomes are lost to an even higher degree than the wild-type upon stress. Cells overexpressing the catalytically inactive rck2-kDa allele are highly sensitive to oxidative stress. In such cells, dissociation of polysomes upon stress was instead greatly delayed. The protein synthesis rate decreased to a similar degree as in wild-type cells, however, indicating that in rck2-kDa cells, the polysome complexes were inactive. Array analyses of total and polysome-associated mRNAs revealed major deregulation of the translational machinery in rck2 mutant cells. This involves transcripts for cytosolic ribosomal proteins and for processing and assembly of ribosomes. In rck2{Delta} cells, weakly transcribed mRNAs associate more avidly with polysomes than in wild-type cells, while the opposite holds true for rck2-kDa cells. This is consistent with perturbed regulation of translation elongation, which is predicted to alter the ratio between mRNAs with and without strong entry sites at ribosomes. We infer that imbalances in the translational apparatus are a major reason for the inability of these cells to respond to stress.

{dagger}Present address: Department of Genetics, Cell Biology, and Development, University of Minnesota at Minneapolis, Minneapolis, MN 55455.

Address correspondence to: Per Sunnerhagen (Per.Sunnerhagen{at}molbio.gu.se)




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