A Variant Family of Mouse Minor Satellite Located on the Centromeric Region of Chromosome 2

Hayashi, Toshimitsu; Ohtsuka, Hiroshi; Kuwabara, Katsuhiro; Mafune, Yoshiro; Miyashita, Nobumoto; Moriwaki, Kazuo; Takahashi, Yoshiaki; Kominami, Ryo

doi:10.1006/geno.1993.1352

Cited by 15 publications

(9 citation statements)

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“…Some aspects of this hierarchical structure may be conserved in M. musculus (32). Two divergent forms of mouse minor satellite that hybridize specifically to chromosome 2 (26) and chromosome 4 (7) have been cloned. This minor satellite, which is otherwise unrelated to ␣-satellite, has a CENP-B binding motif (36).…”

Section: Discussionmentioning

confidence: 99%

Surprising Deficiency of CENP-B Binding Sites in African Green Monkey α-Satellite DNA: Implications for CENP-B Function at Centromeres

Goldberg

Sawhney

Pluta

et al. 1996

Molecular and Cellular Biology

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Centromeres of mammalian chromosomes are rich in repetitive DNAs that are packaged into specialized nucleoprotein structures called heterochromatin. In humans, the major centromeric repetitive DNA, ␣-satellite DNA, has been extensively sequenced and shown to contain binding sites for CENP-B, an 80-kDa centromeric autoantigen. The present report reveals that African green monkey (AGM) cells, which contain extensive ␣-satellite arrays at centromeres, appear to lack the well-characterized CENP-B binding site (the CENP-B box). We show that AGM cells express a functional CENP-B homolog that binds to the CENP-B box and is recognized by several independent anti-CENP-B antibodies. However, three independent assays fail to reveal CENP-B binding sites in AGM DNA. Methods used include a gel mobility shift competition assay using purified AGM ␣-satellite, a novel kinetic electrophoretic mobility shift assay competition protocol using bulk genomic DNA, and bulk sequencing of 76 AGM ␣-satellite monomers. Immunofluorescence studies reveal the presence of significant levels of CENP-B antigen dispersed diffusely throughout the nuclei of interphase cells. These experiments reveal a paradox. CENP-B is highly conserved among mammals, yet its DNA binding site is conserved in human and mouse genomes but not in the AGM genome. One interpretation of these findings is that the role of CENP-B may be in the maintenance and/or organization of centromeric satellite DNA arrays rather than a more direct involvement in centromere structure.

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Section: Discussionmentioning

confidence: 99%

Surprising Deficiency of CENP-B Binding Sites in African Green Monkey α-Satellite DNA: Implications for CENP-B Function at Centromeres

Goldberg

Sawhney

Pluta

et al. 1996

Molecular and Cellular Biology

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“…The size of clones was determined by pulsed-®eld gel electrophoresis as described (Hayashi et al, 1993). Each end of BAC inserts was directly sequenced and PCR primers for the BAC ends were synthesized accordingly.…”

Section: Primer Sequencesmentioning

confidence: 99%

Homozygous deletions and point mutations of the Ikaros gene in γ-ray-induced mouse thymic lymphomas

et al. 1999

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Our previous genome-wide analysis of allelic loss for thymic lymphomas that were induced by g-irradiation in F 1 hybrid mice between BALB/c and MSM strains suggested the centromeric region on chromosome 11 as a site harboring a tumor suppressor gene. Interestingly, to this region the mouse Ikaros gene was mapped which was postulated to participate in oncogenic process from the study of Ikaros knockout mice. Here we show ®ne allelic loss mapping in the vicinity of Ikaros in 191 lymphomas, indicating that the critical region of allelic loss was centered at the Ikaros locus. PCR analysis revealed that nine lymphomas failed to give PCR-ampli®cation for either of two exon primer pairs, indicative of homozygous deletion. Six and ®ve mutations were detected in the N-terminal zinc ®nger domain and the activation domain of Ikaros, respectively, and six of the eleven were frameshift or nonsense mutations that resulted in truncation of Ikaros protein. The results strongly suggest a direct role for Ikaros in development of mouse thymic lymphomas. This provides the experimental basis for further analysis of Ikaros mutations in human cancer.

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“…Pulsed-®eld gel electrophoresis of DNA DNA was prepared in agarose plugs as described (Hayashi et al, 1993). The plugs were digested with a restriction enzyme, NotI, under the manufacturer's recommended conditions, and subjected to electrophoresis using a BioRad She Mapper apparatus.…”

Section: Isolation Of Yacs and Bacs And Analysis Of Their Linkagementioning

confidence: 99%

Allelic loss mapping and physical delineation of a region harboring a putative thymic lymphoma suppressor gene on mouse chromosome 12

et al. 1999

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A Variant Family of Mouse Minor Satellite Located on the Centromeric Region of Chromosome 2

Cited by 15 publications

References 0 publications

Surprising Deficiency of CENP-B Binding Sites in African Green Monkey α-Satellite DNA: Implications for CENP-B Function at Centromeres

Surprising Deficiency of CENP-B Binding Sites in African Green Monkey α-Satellite DNA: Implications for CENP-B Function at Centromeres

Homozygous deletions and point mutations of the Ikaros gene in γ-ray-induced mouse thymic lymphomas

Allelic loss mapping and physical delineation of a region harboring a putative thymic lymphoma suppressor gene on mouse chromosome 12

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