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Genetics, Vol 147, 1025-1041, Copyright © 1997
INVESTIGATIONS |
Mutational Analysis of Cdc19p, a Schizosaccharomyces pombe MCM Protein
S. L. Forsburg, D. A. Sherman, S. Ottilie, J. R. Yasuda and J. A. Hodson
Molecular Biology and Virology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037
The cdc19(+) gene encodes an essential member of the MCM family of replication proteins in Schizosaccharomyces pombe. We have examined the structure and function of the Cdc19p protein using molecular and genetic approaches. We find that overproduction of wild-type Cdc19p in wild-type cells has no effect, but cdc19-P1 mutant cells do not tolerate elevated levels of other MCM proteins or overexpression of mutant forms of Cdc19p. We have found genetic interactions between cdc19(+) and genes encoding subunits of DNA polymerase {delta small} and the replication initiator cdc18(+). We have constructed a series of point mutations and sequence deletions throughout Cdc19p, which allow us to distinguish essential from nonessential regions of the protein. Not surprisingly, conserved residues in the MCM homology domain are required for protein function, but some residues outside the core homology domain are dispensable.
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