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Published March 27, 1987 | public
Journal Article

The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses


Conjugation of ubiquitin to intracellular proteins mediates their selective degradation in eukaryotes. In the yeast Saccharomyces cerevisiae, four distinct ubiquitin-coding loci have been described. UBI1, UBI2, and UBI3 each encode hybrid proteins in which ubiquitin is fused to unrelated sequences. The fourth gene, UBI4, contains five ubiquitin-coding elements in a head-to-tail arrangement, and thus encodes a polyubiquitin precursor protein. A precise, oligonucleotide-directed deletion of UBI4 was constructed in vitro and substituted in the yeast genome in place of the wild-type allele. ubi4 deletion mutants are viable as vegetative cells, grow at wild-type rates, and contain wild-type levels of free ubiquitin under exponential growth conditions. However, although ubi4/UBI4 diploids can form four initially viable spores, the two ubi4 spores within the ascus lose viability extremely rapidly, apparently a novel phenotype in yeast. Furthermore, ubi4/ubi4 diploids are sporulation-defective. ubi4 mutants are also hypersensitive to high temperatures, starvation, and amino acid analogs. These three conditions, while diverse in nature, are all known to induce stress proteins. Expression of the UBI4 gene is similarly induced by either heat stress or starvation. These results indicate that UBI4 is specifically required for the resistance of cells to stress, and that ubiquitin is an essential component of the stress response system.

Additional Information

© 1987 Cell Press. Received 17 September 1986, Revised 5 January 1987. We thank Heung Sik Min for preparing one of the antibodies to ubiquitin, Bonnie Bartel for permission to cite unpublished data, members of David Botstein's laboratory for providing strains, Andrew Murray for demonstrating micromanipulation, members of Jack Szostak's laboratory for allowing use of their micromanipulator, Mark Solomon for comments on the manuscript, Barbara Doran for secretarial assistance, and especially Joan Park for support, advice, and comments on the manuscript. This work was supported by a grant to A. V. from the National Institute of General Medical Sciences (GM31530). E. Ö. was supported by a fellowship from the Charles King Trust and Medical Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 16 USC. Section 1734 solely to indicate this fact.

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