Mouse MCM proteins: complex formation and transportation to the nucleus.
BACKGROUND
The members of the MCM protein family, including MCM2, MCM3, Cdc21, CDC46, Mis5 and CDC47, are considered to be involved in the control of a single round of DNA replication during S phase in eukaryotes. They bind to chromatin during G1 and detach from it during S phase as if they license the chromatin to replicate. However, unlike the originally proposed 'licensing factor' and the budding yeast homologues, mammalian MCM2 and P1MCM3 proteins appeared to be localized in the nucleus during the interphase.


RESULTS
We purified mCdc21 and its associated proteins from mouse cell extract by anti-mCdc21 immunoaffinity chromatography. Three proteins which co-purified with mCdc21 were identified as mCDC47, mMis5 and mMCM2, all were MCM proteins. Glycerol gradient centrifugation analysis showed that all the mouse MCM proteins were detected at 450-600 kDa, an indication of the sum of their calculated molecular weights from their amino acid sequences. mCdc21 was displaced from replicated chromatin in a similar way to P1MCM3 and MCM2 during S phase. Among the six mouse MCM proteins, only mMCM2 and mP1MCM3 showed nuclear localization when overexpressed in COS cells.


CONCLUSIONS
We conclude that in the mouse, six MCM proteins form a single protein complex of molecular weight 450-600 kDa, which may enter the nucleus by nuclear localization signals in the mMCM2 and mP1MCM3 subunits.
Kimura
H
H
Research Centre for Molecular Genetics, Hokkaido University, Kita, Sapporo, Japan.
Ohtomo
T
T
Yamaguchi
M
M
Ishii
A
A
Sugimoto
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001693:Biological Transport, Active; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002501:Centrifugation, Zonal; D002843:Chromatin / Q000378:metabolism; D004261:DNA Replication / Q000502:physiology*; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000302:isolation & purification; D005990:Glycerol; D051379:Mice; D064148:Minichromosome Maintenance Complex Component 4; D064151:Minichromosome Maintenance Complex Component 6; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016196:S Phase; D029702:Schizosaccharomyces pombe Proteins
D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D029702:Schizosaccharomyces pombe Proteins; D064148:Minichromosome Maintenance Complex Component 4; D064151:Minichromosome Maintenance Complex Component 6; C078490:mcm4 protein, S pombe; C092300:mcm6 protein, S pombe; D005990:Glycerol