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Molecular and Cellular Biology, April 2000, p. 2852-2864, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Fission Yeast Homologs of Human CENP-B Have Redundant Functions Affecting Cell Growth and Chromosome Segregation

Mary Baum and Louise Clarke*

Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, California 93106

Received 30 June 1999/Returned for modification 26 August 1999/Accepted 14 January 2000

Two functionally important DNA sequence elements in centromeres of the fission yeast Schizosaccharomyces pombe are the centromeric central core and the K-type repeat. Both of these DNA elements show internal functional redundancy that is not correlated with a conserved DNA sequence. Specific, but degenerate, sequences in these elements are bound in vitro by the S. pombe DNA-binding proteins Abp1p (also called Cbp1p) and Cbhp, which are related to the mammalian centromere DNA-binding protein CENP-B. In this study, we determined that Abp1p binds to at least one of its target sequences within S. pombe centromere II central core (cc2) DNA with an affinity (Ks = 7 × 109 M-1) higher than those of other known centromere DNA-binding proteins for their cognate targets. In vivo, epitope-tagged Cbhp associated with centromeric K repeat chromatin, as well as with noncentromeric regions. Like abp1+/cbp1+, we found that cbh+ is not essential in fission yeast, but a strain carrying deletions of both genes (Delta abp1 Delta cbh) is extremely compromised in growth rate and morphology and missegregates chromosomes at very high frequency. The synergism between the two null mutations suggests that these proteins perform redundant functions in S. pombe chromosome segregation. In vitro assays with cell extracts with these proteins depleted allowed the specific assignments of several binding sites for them within cc2 and the K-type repeat. Redundancy observed at the centromere DNA level appears to be reflected at the protein level, as no single member of the CENP-B-related protein family is essential for proper chromosome segregation in fission yeast. The relevance of these findings to mammalian centromeres is discussed.

* Corresponding author. Mailing address: Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA 93106. Phone: (805) 893-3624. Fax: (805) 893-4724. E-mail: clarke{at}lifesci.lscf.ucsb.edu.

Molecular and Cellular Biology, April 2000, p. 2852-2864, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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