Centromere Protein B Null Mice are Mitotically and Meiotically Normal but Have Lower Body and Testis Weights

CENP-B is a constitutive centromere DNA-binding protein that is conserved in a number of mammalian species and in yeast. Despite this conservation, earlier cytological and indirect experimental studies have provided conflicting evidence concerning the role of this protein in mitosis. The requirement of this protein in meiosis has also not previously been described. To resolve these uncertainties, we used targeted disruption of the Cenpb gene in mouse to study the functional significance of this protein in mitosis and meiosis. Male and female Cenpb null mice have normal body weights at birth and at weaning, but these subsequently lag behind those of the heterozygous and wild-type animals. The weight and sperm content of the testes of Cenpb null mice are also significantly decreased. Otherwise, the animals appear developmentally and reproductively normal. Cytogenetic fluorescence-activated cell sorting and histological analyses of somatic and germline tissues revealed no abnormality. These results indicate that Cenpb is not essential for mitosis or meiosis, although the observed weight reduction raises the possibility that Cenpb deficiency may subtly affect some aspects of centromere assembly and function, and result in reduced rate of cell cycle progression, efficiency of microtubule capture, and/or chromosome movement. A model for a functional redundancy of this protein is presented.

[1]  M. Vidal,et al.  Identification of a family of human F-box proteins , 1999, Current Biology.

[2]  K. Choo,et al.  Targeted disruption of mouse centromere protein C gene leads to mitotic disarray and early embryo death. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[3]  G. Karpen,et al.  The case for epigenetic effects on centromere identity and function. , 1997, Trends in genetics : TIG.

[4]  K. Choo Centromere DNA dynamics: latent centromeres and neocentromere formation. , 1997, American journal of human genetics.

[5]  W. Brown,et al.  Efficient conditional mutation of the vertebrate CENP-C gene. , 1997, Human molecular genetics.

[6]  J. Hurwitz,et al.  Purification and characterization of a CENP-B homologue protein that binds to the centromeric K-type repeat DNA of Schizosaccharomyces pombe. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Schwartz,et al.  Characterization of neo-centromeres in marker chromosomes lacking detectable alpha-satellite DNA. , 1997, Human molecular genetics.

[8]  P. Warburton,et al.  Centromeres, CENP-B and Tigger too. , 1997, Trends in genetics : TIG.

[9]  M. Baum,et al.  A Centromere DNA-binding Protein from Fission Yeast Affects Chromosome Segregation and Has Homology to Human CENP-B , 1997, The Journal of cell biology.

[10]  W. Earnshaw,et al.  Surprising deficiency of CENP-B binding sites in African green monkey alpha-satellite DNA: implications for CENP-B function at centromeres , 1996, Molecular and cellular biology.

[11]  H. Masumoto,et al.  Centromere protein B of African green monkey cells: gene structure, cellular expression, and centromeric localization , 1996, Molecular and cellular biology.

[12]  A. Mushegian,et al.  Evidence for selection in evolution of alpha satellite DNA: the central role of CENP-B/pJ alpha binding region. , 1996, Journal of molecular biology.

[13]  J. Sedat,et al.  Direct Evidence of a Role for Heterochromatin in Meiotic Chromosome Segregation , 1996, Cell.

[14]  G. Karpen,et al.  Centric Heterochromatin and the Efficiency of Achiasmate Disjunction in Drosophila Female Meiosis , 1996, Science.

[15]  M. Valdivia,et al.  Molecular cloning of an intronless gene for the hamster centromere antigen CENP-B. , 1996, Biochimica et biophysica acta.

[16]  W. Earnshaw,et al.  The Centromere: Hub of Chromosomal Activities , 1995, Science.

[17]  S. Schwartz,et al.  Identification of centromeric antigens in dicentric Robertsonian translocations: CENP-C and CENP-E are necessary components of functional centromeres. , 1995, Human molecular genetics.

[18]  N. Wreford,et al.  Theory and practice of stereological techniques applied to the estimation of cell number and nuclear volume in the testis , 1995, Microscopy research and technique.

[19]  H. Masumoto,et al.  CENP-B binds a novel centromeric sequence in the Asian mouse Mus caroli , 1995, Molecular and cellular biology.

[20]  M. T. Brown,et al.  Sequence similarities between the yeast chromosome segregation protein Mif2 and the mammalian centromere protein CENP-C. , 1995, Gene.

[21]  D. Koshland,et al.  Evidence that the MIF2 gene of Saccharomyces cerevisiae encodes a centromere protein with homology to the mammalian centromere protein CENP-C. , 1995, Molecular biology of the cell.

[22]  H. Masumoto,et al.  Analysis of protein-DNA and protein-protein interactions of centromere protein B (CENP-B) and properties of the DNA-CENP-B complex in the cell cycle , 1995, Molecular and cellular biology.

[23]  S. Stoler,et al.  A mutation in CSE4, an essential gene encoding a novel chromatin-associated protein in yeast, causes chromosome nondisjunction and cell cycle arrest at mitosis. , 1995, Genes & development.

[24]  L. Shaffer,et al.  Further evidence that CENP-C is a necessary component of active centromeres: studies of a dic(X; 15) with simultaneous immunofluorescence and FISH. , 1995, Human molecular genetics.

[25]  Y. Fukushima,et al.  A stable acentric marker chromosome: possible existence of an intercalary ancient centromere at distal 8p. , 1994, American journal of human genetics.

[26]  J. Mann,et al.  Quantitative RT-PCR assays show Xist RNA levels are low in mouse female adult tissue, embryos and embryoid bodies. , 1994, Development.

[27]  K. Sullivan,et al.  Human CENP-A contains a histone H3 related histone fold domain that is required for targeting to the centromere , 1994, The Journal of cell biology.

[28]  T. Saito,et al.  Mapping of the human centromere protein B gene (CENPB) to chromosome 20p13 by fluorescence in situ hybridization. , 1994, Genomics.

[29]  W. Earnshaw,et al.  Role of nonhistone proteins in the chromosomal events of mitosis , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[30]  H. Masumoto,et al.  Distribution of CENP-B boxes reflected in CREST centromere antigenic sites on long-range alpha-satellite DNA arrays of human chromosome 21. , 1994, Human molecular genetics.

[31]  J. Nichols,et al.  Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[32]  K. Choo,et al.  A novel nuclear protein binds centromeric alpha satellite DNA. , 1994, Human molecular genetics.

[33]  W. Earnshaw,et al.  CENP-C is required for maintaining proper kinetochore size and for a timely transition to anaphase , 1994, The Journal of cell biology.

[34]  R. Durbin,et al.  2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans , 1994, Nature.

[35]  L. Hartwell,et al.  MIF2 is required for mitotic spindle integrity during anaphase spindle elongation in Saccharomyces cerevisiae , 1993, The Journal of cell biology.

[36]  B. Vissel,et al.  Long-range analyses of the centromeric regions of human chromosomes 13, 14 and 21: identification of a narrow domain containing two key centromeric DNA elements. , 1993, Human molecular genetics.

[37]  J. Roder,et al.  Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Sugimoto,et al.  Mapping of the human CENP-B gene to chromosome 20 and the CENP-C gene to chromosome 12 by a rapid cycle DNA amplification procedure. , 1993, Genomics.

[39]  L. Voullaire,et al.  A functional marker centromere with no detectable alpha-satellite, satellite III, or CENP-B protein: activation of a latent centromere? , 1993, American journal of human genetics.

[40]  L. Romanova,et al.  Definition of a new alpha satellite suprachromosomal family characterized by monomeric organization. , 1993, Nucleic acids research.

[41]  H. Masumoto,et al.  A human centromere protein, CENP-B, has a DNA binding domain containing four potential alpha helices at the NH2 terminus, which is separable from dimerizing activity , 1992, The Journal of cell biology.

[42]  B. Schaar,et al.  CENP-E is a putative kinetochore motor that accumulates just before mitosis , 1992, Nature.

[43]  W. Earnshaw,et al.  CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate , 1992, Cell.

[44]  C. Jones,et al.  Higher rate of evolution of X chromosome alpha‐repeat DNA in human than in the great apes. , 1992, The EMBO journal.

[45]  W. Earnshaw,et al.  Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere , 1992, The Journal of cell biology.

[46]  K. Yoda,et al.  Centromere protein B assembles human centromeric alpha-satellite DNA at the 17-bp sequence, CENP-B box , 1992, The Journal of cell biology.

[47]  W. Earnshaw,et al.  Disruption of centromere assembly during interphase inhibits kinetochore morphogenesis and function in mitosis , 1991, Cell.

[48]  R. Zinkowski,et al.  CENP‐E, a novel human centromere‐associated protein required for progression from metaphase to anaphase. , 1991, The EMBO journal.

[49]  B. Vissel,et al.  A SURVEY OF THE GENOMIC DISTRIBUTION OF ALPHA SATELLITE DNA ON ALL THE HUMAN CHROMOSOMES, AND DERIVATION OF A NEW CONSENSUS SEQUENCE , 1991 .

[50]  J. B. Rattner,et al.  The structure of the mammalian centromere , 1991, BioEssays : news and reviews in molecular, cellular and developmental biology.

[51]  G. Borisy,et al.  Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis , 1990, The Journal of cell biology.

[52]  G. Schatten,et al.  Microinjected centromere [corrected] kinetochore antibodies interfere with chromosome movement in meiotic and mitotic mouse oocytes [published erratum appears in J Cell Biol 1990 Dec;111(6 Pt 1):following 2800] , 1990, The Journal of cell biology.

[53]  W. Earnshaw,et al.  CENP-B: a major human centromere protein located beneath the kinetochore , 1990, The Journal of cell biology.

[54]  C. Murphy,et al.  Satellite 1 DNA sequence from genomic DNA of the giant panda Ailuropoda melanoleuca. , 1990, Nucleic acids research.

[55]  H. Masumoto,et al.  Alphoid satellite DNA is tightly associated with centromere antigens in human chromosomes throughout the cell cycle. , 1989, Experimental cell research.

[56]  Lukas C. Kühn,et al.  A stem-loop in the 3′ untranslated region mediates iron-dependent regulation of transferrin receptor mRNA stability in the cytoplasm , 1988, Cell.

[57]  C. Tyler-Smith,et al.  Structure of the major block of alphoid satellite DNA on the human Y chromosome. , 1987, Journal of molecular biology.

[58]  T. Pollard,et al.  Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen , 1987, The Journal of cell biology.

[59]  R. Margolis,et al.  A 17-kD centromere protein (CENP-A) copurifies with nucleosome core particles and with histones , 1987, The Journal of cell biology.

[60]  A. Cerami,et al.  Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[61]  V. Chapman,et al.  Construction of a small Mus musculus repetitive DNA library: identification of a new satellite sequence in Mus musculus. , 1983, Nucleic acids research.

[62]  E. Tan,et al.  Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients , 1981, The Journal of cell biology.

[63]  E. Tan,et al.  Distribution of kinetochore (centromere) antigen in mammalian cell nuclei , 1981, The Journal of cell biology.

[64]  M. Fritzler,et al.  The CREST syndrome: a distinct serologic entity with anticentromere antibodies. , 1980, The American journal of medicine.

[65]  R. P. Amann,et al.  Daily sperm production and epididymal sperm reserves of pubertal and adult rats. , 1978, Journal of reproduction and fertility.

[66]  K. Choo,et al.  A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DNA , 1997, Nature Genetics.

[67]  D. Ward,et al.  Rabl orientation of CENP-B box sequences in Tupaia belangeri fibroblasts. , 1995, Cytogenetics and cell genetics.

[68]  R. Zinkowski,et al.  Structure and molecular organization of the centromere-kinetochore complex. , 1992, Trends in cell biology.

[69]  B. Vissel,et al.  A survey of the genomic distribution of alpha satellite DNA on all the human chromosomes, and derivation of a new consensus sequence. , 1991, Nucleic acids research.

[70]  R. Margolis,et al.  Kinetochore components recognized by human autoantibodies are present on mononucleosomes. , 1985, Molecular and cellular biology.

[71]  J. Erickson What Is the Centromere , 1979 .