Mts4, a Non-ATPase Subunit of the 26 S Protease in Fission Yeast Is Essential for Mitosis and Interacts Directly with the ATPase Subunit Mts2*
- From the ‡MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh, EH4 2XU Scotland, United Kingdom and the ¶Institute of Biochemistry, Medical Faculty, Humboldt University, Monbijoustrasse 2A 10117, Berlin, Federal Republic of Germany
Abstract
We have isolated a fission yeast gene,mts4 +, by complementation of a temperature-sensitive mutation and show that it encodes subunit 2 (S2) of the 19 S regulatory complex of the 26 S protease.mts4 + is an essential gene, and we show that loss of this subunit causes cells to arrest in metaphase, illustrating the importance of S2 for mitosis. The Mts4 protein is 48% identical to S2 of the human 26 S protease, and the lethal phenotype of the nullmts4 allele can be rescued by the human cDNA encoding S2. We provide genetic and physical evidence to suggest that the Mts4 protein interacts with the product of the mts2 +gene, an ATPase which has previously been shown to be subunit 4 of the 26 S protease.
Footnotes
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↵* This work was supported by Medical Research Council funding (to C. W. and C. G.), a Boehringer Ingelheim short term fellowship (to M. S.), and by a research grant from the Deutsche Forschungsgemeinschaft (to W. D.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .
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↵§ To whom correspondence should be addressed. Tel.: 131 332 2471; Fax: 131 343 2620.
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↵1 The abbreviations used are: PAGE, polyacrylamide gel electrophoresis; PCR, polymerase chain reaction; GST, glutathioneS-transferase; DAPI, 4′,6-diamidino-2-phenylindole; ts, temperature-sensitive; TNF, tumor necrosis factor; MBC, methyl benzimidazol-2-yl carbamylate; APC, anaphase promoting complex; ts, temperature-sensitive.
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↵2 C. R. M. Wilkinson and C. Gordon, unpublished data.
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- Received January 30, 1997.
- Revision received June 23, 1997.
