Nucleic Acids Research, 2000, Vol. 28, No. 3 809-817
© 2000 Oxford University Press
Isolation and characterization of human orthologs of yeast CCR4–NOT complex subunits
Laboratory for Physiological Chemistry and Centre for Biomedical Genetics, Utrecht University, PO Box 80042, 3508 TA Utrecht, The Netherlands, 1Department of Medical Biochemistry, University of Geneva Medical School, CMU, 1 rue Michel Servet, 1211 Geneva 4, Switzerland and 2Human Genome Sciences, Inc., 9410 Key West Avenue, Rockville, MD 20850, USA
The yeast CCR4–NOT protein complex is a global regulator of RNA polymerase II transcription. It is comprised of yeast NOT1 to NOT5, yeast CCR4 and additional proteins like yeast CAF1. Here we report the isolation of cDNAs encoding human NOT2, NOT3, NOT4 and a CAF1-like factor, CALIF. Analysis of their mRNA levels in different human tissues reveals a common ubiquitous expression pattern. A multitude of two-hybrid interactions among the human cDNAs suggest that their encoded proteins also form a complex in mammalian cells. Functional conservation of these proteins throughout evolution is supported by the observation that the isolated human NOT3 and NOT4 cDNAs can partially complement corresponding not mutations in yeast. Interestingly, human CALIF is highly homologous to, although clearly different from, a recently described human CAF1 protein. Conserved interactions of this factor with both NOT and CCR4 proteins and co-immunoprecipitation experiments suggest that CALIF is a bona fide component of the human CCR4–NOT complex.
* To whom correspondence should be addressed. Tel: +31 30 2538981; Fax: +31 30 2539035; Email: h.t.m.timmers@med.uu.nl
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