Nucleic Acids Research, 1992, Vol. 20, No. 5 1069-1074
© 1992
MOLECULAR BIOLOGY |
Properties of the nuclear P1 protein, a mammalian homologue of the yeast Mcm3 replication protein
Division of Biology, Universitat Konstanz D775 Konstanz, FRG , USA 1Department of Pharmacology, University of Massachusetts Medical School Worcester, MA 01605, USA
*To whom correspondence should be addressed
Received December 4, 1991. Revised February 6, 1992. Accepted February 6, 1992.
Polyclonal antibodies were raised against a multiprotein ‘holoenzyme’ form of calf thymus DNA polymerase 
-primase and used to probe a human cDNA-protein expression library constructed in the 
gt11 vector. The probe identified a series of cDNA clones derived from a 3.2 kb mRNA which encodes a novel 105 kDa polypeptide, the P1 protein. In Intact cells, the P1 protein was specifically associated with the nucleus, and in cell extracts, it was associated with complex forms of DNA polymerase -prlmase. The synthesis of human P1-specific mRNA was stimulated upon addition of fresh serum to growth-arrested cells, and RNA blot analyses with the human PI-cDNA probe indicated that P1 is encoded by a strictly conserved mammalian gene. The amino acid sequence deduced from a 240-codon open reading frame resident in the largest human PI-cDNA (0.84 kb) displayed >96% identity with that deduced from the equivalent segment of a 795-codon open reading frame of a larger mouse PI-cDNA (2.8 kb). Throughout its length, the primary structure of mammalian P1 displayed strong homology with that of Mcm3, a 125 kDa yeast protein thought to be involved in the initiation of DNA replication (Gibson et al. 1990. Mol. Cell. Biol. 10: 5707–5720). The P1-Mcm3 homology, the strong conservation of P1 among mammals, its nuclear localization, and Its association with the replication-specific DNA polymerase strongly suggest an important role of the P1 protein in the replication of mammalian DNA.
+Present addresses: Department of Biology, Imperial College, Prince Consort Road, London, UK
Present addresses: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA
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