Journal of Cell Science, Vol 109, Issue 2 319-326, Copyright © 1996 by Company of Biologists

JOURNAL ARTICLES

Fission yeast Nda1 and Nda4, MCM homologs required for DNA replication, are constitutive nuclear proteins

N Okishio, Y Adachi and M Yanagida
Department of Biophysics, Kyoto University, Japan.

The nda1+ and nda4+ genes of the fission yeast Schizosaccharomyces pombe encode proteins similar to budding yeast MCM2 and MCM5/CDC46, respectively, which are required for the early stages of DNA replication. The budding yeast Mcm proteins display cell-cycle dependent localization. They are present in the nucleus specifically from late M phase until the beginning of S phase, so that they were suggested to be components of a replication licensing factor, a positive factor for the onset of replication, which is thought to be inactivated after use, thus restricting replication to only once in a cell cycle. In the present study, we raised antibodies against Nda1 or Nda4 and identified 115 kDa and 80 kDa proteins, respectively. Their immunolocalization was examined in wild-type cells and in various cell-cycle mutants. Both Nda1 and Nda4 proteins remained primarily in the nucleus throughout the cell cycle. In mutants arrested in G1, S, and G2 phases, these proteins were also enriched in the nucleus. These results indicate that the dramatic change in subcellular localization as seen in budding yeast is not essential in fission yeast for the functions of Nda1 and Nda4 proteins to be executed. The histidine-tagged nda1+ gene was constructed and integrated into the chromosome to replace the wild-type nda1+ gene. The resulting His-tagged Nda1 protein was adsorbed to the Ni-affinity column, and co-eluted with the untagged Nda4 protein, suggesting that they formed a complex.

This article has been cited by other articles:

				T. Tvegard, H. Soltani, H. C. Skjolberg, M. Krohn, E. A. Nilssen, S. E. Kearsey, B. Grallert, and E. Boye A novel checkpoint mechanism regulating the G1/S transition Genes & Dev., March 15, 2007; 21(6): 649 - 654. [Abstract] [Full Text] [PDF]

				S. L. Forsburg Eukaryotic MCM Proteins: Beyond Replication Initiation Microbiol. Mol. Biol. Rev., March 1, 2004; 68(1): 109 - 131. [Abstract] [Full Text] [PDF]

				J. A. Hodson, J. M. Bailis, and S. L. Forsburg Efficient labeling of fission yeast Schizosaccharomyces pombe with thymidine and BUdR Nucleic Acids Res., November 1, 2003; 31(21): e134 - e134. [Abstract] [Full Text] [PDF]

				E. B. Gomez, M. G. Catlett, and S. L. Forsburg Different Phenotypes in Vivo Are Associated With ATPase Motif Mutations in Schizosaccharomyces pombe Minichromosome Maintenance Proteins Genetics, April 1, 2002; 160(4): 1305 - 1318. [Abstract] [Full Text] [PDF]

				D. T. Liang and S. L. Forsburg Characterization of Schizosaccharomyces pombe mcm7+ and cdc23+ (MCM10) and Interactions With Replication Checkpoints Genetics, October 1, 2001; 159(2): 471 - 486. [Abstract] [Full Text] [PDF]

				A. Vas, W. Mok, and J. Leatherwood Control of DNA Rereplication via Cdc2 Phosphorylation Sites in the Origin Recognition Complex Mol. Cell. Biol., September 1, 2001; 21(17): 5767 - 5777. [Abstract] [Full Text] [PDF]

				H. A. Snaith, G. W. Brown, and S. L. Forsburg Schizosaccharomyces pombe Hsk1p Is a Potential Cds1p Target Required for Genome Integrity Mol. Cell. Biol., November 1, 2000; 20(21): 7922 - 7932. [Abstract] [Full Text]

				S. G. Pasion and S. L. Forsburg Nuclear Localization of Schizosaccharomyces pombe Mcm2/Cdc19p Requires MCM Complex Assembly Mol. Biol. Cell, December 1, 1999; 10(12): 4043 - 4057. [Abstract] [Full Text]

				Y. Ogawa, T. Takahashi, and H. Masukata Association of Fission Yeast Orp1 and Mcm6 Proteins with Chromosomal Replication Origins Mol. Cell. Biol., October 1, 1999; 19(10): 7228 - 7236. [Abstract] [Full Text] [PDF]

				H. A. Snaith and S. L. Forsburg Rereplication Phenomenon in Fission Yeast Requires MCM Proteins and Other S Phase Genes Genetics, July 1, 1999; 152(3): 839 - 851. [Abstract] [Full Text]

				D. Liang, J. Hodson, and S. Forsburg Reduced dosage of a single fission yeast MCM protein causes genetic instability and S phase delay J. Cell Sci., January 2, 1999; 112(4): 559 - 567. [Abstract] [PDF]

				K. L. Gould, C. G. Burns, A. Feoktistova, C.-P. Hu, S. G. Pasion, and S. L. Forsburg Fission Yeast cdc24+ Encodes a Novel Replication Factor Required for Chromosome Integrity Genetics, July 1, 1998; 149(3): 1221 - 1233. [Abstract] [Full Text] [PDF]

				D. A. Sherman, S. G. Pasion, and S. L. Forsburg Multiple Domains of Fission Yeast Cdc19p (MCM2) Are Required for Its Association with the Core MCM Complex Mol. Biol. Cell, July 1, 1998; 9(7): 1833 - 1845. [Abstract] [Full Text]

				X. S. Ye, R. R. Fincher, A. Tang, K. K. McNeal, S. E. Gygax, A. N. Wexler, K. B. Ryan, S. W. James, and S. A. Osmani Proteolysis and Tyrosine Phosphorylation of p34cdc2/Cyclin B. THE ROLE OF MCM2 AND INITIATION OF DNA REPLICATION TO ALLOW TYROSINE PHOSPHORYLATION OF p34cdc2 J. Biol. Chem., December 26, 1997; 272(52): 33384 - 33393. [Abstract] [Full Text] [PDF]

				K Shimada, M Harata, and S Mizuno A nuclear matrix-associated high molecular mass nuclear antigen, HMNA, of chicken and marked decrease of its immunoreactivity during the progression of S phase J. Cell Sci., January 12, 1997; 110(24): 3031 - 3041. [Abstract] [PDF]

				D. F. Shechter, C. Y. Ying, and J. Gautier The Intrinsic DNA Helicase Activity of Methanobacterium thermoautotrophicum Delta H Minichromosome Maintenance Protein J. Biol. Chem., May 12, 2000; 275(20): 15049 - 15059. [Abstract] [Full Text] [PDF]

				J.-K. Lee and J. Hurwitz Isolation and Characterization of Various Complexes of the Minichromosome Maintenance Proteins of Schizosaccharomyces pombe J. Biol. Chem., June 16, 2000; 275(25): 18871 - 18878. [Abstract] [Full Text] [PDF]

				M. Uchiyama, D. Griffiths, K.-i. Arai, and H. Masai Essential Role of Sna41/Cdc45 in Loading of DNA Polymerase alpha onto Minichromosome Maintenance Proteins in Fission Yeast J. Biol. Chem., July 6, 2001; 276(28): 26189 - 26196. [Abstract] [Full Text] [PDF]