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J. Biol. Chem., Vol. 277, Issue 4, 2835-2842, January 25, 2002
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and
From the Department of Medical Biochemistry, University of
Geneva Medical School, 1 rue Michel Servet,
1211 Geneva 4, Switzerland
The Ccr4-Not complex is a global regulator of
transcription that affects genes positively and negatively and is
thought to regulate transcription factor IID function. Two components
of this complex, Caf1p and Ccr4p, are directly involved in mRNA
deadenylation, and Caf1p is associated with Dhh1p, a putative RNA
helicase thought to be a component of the decapping complex. In this
work, we tried to determine whether Dhh1p might interact with the
Ccr4-Not complex. We found that, first, not mutations
displayed severe synthetic phenotypes when combined with a
dhh1-null mutation. Second, overexpression of Not1p was
toxic in dhh1-null cells. Third, a not mutant
phenotype was suppressed by deletion of DHH1 and mimicked
by overexpression of DHH1. Fourth, dhh1-null
mutants displayed resistance to heat shock, a phenotype observed for
all mutants that affect the Ccr4-Not complex. Finally, like Caf1p and
Ccr4p, Dhh1p co-immunoprecipitated with the nonessential N-terminal
domain of Not1p, and the levels of Caf1p and Dhh1p were dependent upon
this Not1p domain. Taken together, our results suggest that the
Ccr4-Not complex, via the N-terminal region of Not1p, is necessary for
the maintenance of stable cellular levels of Dhh1p and Caf1p, thus
contributing to regulation of mRNA decay in addition to transcription.
Present address: Inst. de Biochimie et Génétique
Cellulaires CNRS-UMR5095, 1 rue Camille Saint-Saens, 33077 Bordeaux
Cedex, France.
§
To whom correspondence should be addressed. Tel.: 41-22-702-54-76;
Fax: 41-22-702-55-02; E-mail: martine.collart@medecine.unige.ch.
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