A human nuclear protein with sequence homology to a family of early S phase proteins is required for entry into S phase and for cell division

J Cell Sci. 1994 Jan:107 ( Pt 1):253-65. doi: 10.1242/jcs.107.1.253.

Abstract

Molecular cloning and characterisation of a human nuclear protein designated BM28 is reported. On the amino acid level this 892 amino acid protein, migrating on SDS-gels as a 125 kDa polypeptide, shares areas of significant similarity with a recently defined family of early S phase proteins. The members of this family, the Saccharomyces cerevisiae Mcm2p, Mcm3p, Cdc46p/Mcm5p, the Schizosaccharomyces pombe Cdc21p and the mouse protein P1 are considered to be involved in the onset of DNA replication. The highest similarity was found with Mcm2p (42% identity over the whole length and higher than 75% over a conservative region of 215 amino acid residues), suggesting that BM28 could represent the human homologue of the S. cerevisiae MCM2. Using antibodies raised against the recombinant BM28 the corresponding antigen was found to be localised in the nuclei of various mammalian cells. Microinjection of anti-BM28 antibody into synchronised mouse NIH3T3 or human HeLa cells presents evidence for the involvement of the protein in cell cycle progression. When injected in G1 phase the anti-BM28 antibody inhibits the onset of subsequent DNA synthesis as tested by the incorporation of bromodeoxyuridine. Microinjection during the S phase had no effect on DNA synthesis, but inhibits cell division. The data suggest that the nuclear protein BM28 is required for two events of the cell cycle, for the onset of DNA replication and for cell division.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Antibodies / pharmacology
  • Cell Cycle Proteins*
  • Cell Division*
  • Cell Line
  • Cloning, Molecular
  • Consensus Sequence
  • DNA / biosynthesis
  • DNA Replication
  • HeLa Cells
  • Humans
  • Kinetics
  • Mice
  • Minichromosome Maintenance Complex Component 2
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • S Phase*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism
  • Sequence Homology, Amino Acid

Substances

  • Antibodies
  • Cell Cycle Proteins
  • Nuclear Proteins
  • DNA
  • MCM2 protein, human
  • Minichromosome Maintenance Complex Component 2

Associated data

  • GENBANK/P03029
  • GENBANK/P06519
  • GENBANK/P07604
  • GENBANK/P13632
  • GENBANK/P17899
  • GENBANK/P19323
  • GENBANK/P23737
  • GENBANK/P24279
  • GENBANK/P25206
  • GENBANK/P29458
  • GENBANK/P29469
  • GENBANK/P29496
  • GENBANK/S06965
  • GENBANK/X67334
  • GENBANK/Z21487