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Vol. 9, Issue 7, 1833-1845, July 1998
*Molecular Biology and Virology Laboratory, The Salk Institute, La
Jolla, California 92037; and
The members of the MCM protein family are essential eukaryotic DNA
replication factors that form a six-member protein complex. In this
study, we use antibodies to four MCM proteins to investigate the
structure of and requirements for the formation of fission yeast MCM
complexes in vivo, with particular regard to Cdc19p (MCM2). Gel
filtration analysis shows that the MCM protein complexes are unstable
and can be broken down to subcomplexes. Using coimmunoprecipitation, we
find that Mis5p (MCM6) and Cdc21p (MCM4) are tightly associated with
one another in a core complex with which Cdc19p loosely associates. Assembly of Cdc19p with the core depends upon Cdc21p. Interestingly, there is no obvious change in Cdc19p-containing MCM complexes through
the cell cycle. Using a panel of Cdc19p mutants, we find that multiple
domains of Cdc19p are required for MCM binding. These studies indicate
that MCM complexes in fission yeast have distinct substructures, which
may be relevant for function.
Department of Biology,
University of California, San Diego, La Jolla, California 92093-0348
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