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Molecular and Cellular Biology, October 2000, p. 7080-7087, Vol. 20, No. 19
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
BUR1 and BUR2 Encode a Divergent
Cyclin-Dependent Kinase-Cyclin Complex Important for Transcription
In Vivo
Sheng
Yao,1
Aaron
Neiman,2 and
Gregory
Prelich1,*
Department of Molecular Genetics, Albert
Einstein College of Medicine, Bronx, New York
10461,1 and Department of
Biochemistry and Cell Biology, State University of New York at
Stony Brook, Stony Brook, New York 11794-52152
Received 9 May 2000/Returned for modification 12 June 2000/Accepted 11 July 2000
BUR1 and BUR2 were previously identified by
a genetic selection for mutations that increase transcription from
basal promoters in vivo. BUR1 encoded a putative protein
kinase with greatest similarity to members of the cyclin-dependent
kinase (CDK) family, although that similarity was not sufficient to
classify it as a CDK. It was also not known whether Bur1 activity was
cyclin dependent and, if so, which cyclins stimulated Bur1. The
molecular cloning and characterization of BUR2 presented
here sheds light on these issues. Genetic analysis indicates that
BUR2 function is intimately related to that of
BUR1: bur1 and bur2 mutations cause
nearly identical spectra of mutant phenotypes, and overexpression of
BUR1 suppresses a bur2 null allele. Biochemical
analysis has provided a molecular basis for these genetic observations.
We find that BUR2 encodes a cyclin for the Bur1 protein
kinase, based on the following evidence. First, the BUR2
amino acid sequence reveals similarity to the cyclins; second, Bur1 and
Bur2 coimmunoprecipitate from crude extracts and interact in the
two-hybrid system; and third, BUR2 is required for Bur1
kinase activity in vitro. Our combined genetic and biochemical results
therefore indicate that Bur1 and Bur2 comprise a divergent CDK-cyclin
complex that has an important functional role during transcription in vivo.
*
Corresponding author. Mailing address: Department of
Molecular Genetics, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-2181. Fax: (718) 430-8778. E-mail: prelich{at}aecom.yu.edu.
Molecular and Cellular Biology, October 2000, p. 7080-7087, Vol. 20, No. 19
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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