Centromere protein B of African green monkey cells

Gene structure, cellular expression, and centromeric localization

Kinya Yoda, Takao Nakamura, Hiroshi Masumoto, Nobutaka Suzuki, Katsumi Kitagawa, Megumi Nakano, Akihisa Shinjo, Tuneko Okazaki

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Centromere protein B (CENP-B) is a centromeric DNA-binding protein which recognizes a 17-bp sequence (CENP-B box) in human and mouse centromeric satellite DNA. The African green monkey (AGM) is phylogenetically closer to humans than mice and is known to contain large amounts of α-satellite DNA, but there has been no report of CENP-B boxes or CENP-B in the centromere domains of its chromosomes. To elucidate the AGM CENP-B-CENP-B box interaction, we have analyzed the gene structure, expression, biochemical properties, and centromeric localization of its CENP-B. The amino acid sequence deduced from the cloned AGM CENP-B gene was established to be highly homologous to that of human and mouse CENP-B. In particular, the DNA binding and homodimer formation domains demonstrated 100% identity to their human and mouse counterparts. Immunoblotting and DNA mobility shift analyses revealed CENP-B to be expressed in AGM cell lines. As predicted from the gene structure, the AGM CENP-B in the cell extracts exhibited the same DNA binding specificity and homodimer forming activity as human CENP-B. By indirect immunofluorescent staining of AGM mitotic cells with anti-CENP-B antibodies, a centromere-specific localization of AGM CENP- B could be demonstrated. We also isolated AGM α-satellite DNA with a CENP- B box-like sequence with CENP-B affinity. These results not only prove that CENP-B functionally persists in AGM cells but also suggest that the AGM genome contains the recognition sequences for CENP-B (CENP-B boxes with the core recognition sequence or CENP-B box variants) in centromeric satellite DNA.

Original languageEnglish (US)
Pages (from-to)5169-5177
Number of pages9
JournalMolecular and Cellular Biology
Volume16
Issue number9
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

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Centromere Protein B
Cercopithecus aethiops
Cellular Structures
Genes
Satellite DNA
Centromere
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Centromere protein B of African green monkey cells : Gene structure, cellular expression, and centromeric localization. / Yoda, Kinya; Nakamura, Takao; Masumoto, Hiroshi; Suzuki, Nobutaka; Kitagawa, Katsumi; Nakano, Megumi; Shinjo, Akihisa; Okazaki, Tuneko.

In: Molecular and Cellular Biology, Vol. 16, No. 9, 01.01.1996, p. 5169-5177.

Research output: Contribution to journalArticle

Yoda, Kinya ; Nakamura, Takao ; Masumoto, Hiroshi ; Suzuki, Nobutaka ; Kitagawa, Katsumi ; Nakano, Megumi ; Shinjo, Akihisa ; Okazaki, Tuneko. / Centromere protein B of African green monkey cells : Gene structure, cellular expression, and centromeric localization. In: Molecular and Cellular Biology. 1996 ; Vol. 16, No. 9. pp. 5169-5177.
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abstract = "Centromere protein B (CENP-B) is a centromeric DNA-binding protein which recognizes a 17-bp sequence (CENP-B box) in human and mouse centromeric satellite DNA. The African green monkey (AGM) is phylogenetically closer to humans than mice and is known to contain large amounts of α-satellite DNA, but there has been no report of CENP-B boxes or CENP-B in the centromere domains of its chromosomes. To elucidate the AGM CENP-B-CENP-B box interaction, we have analyzed the gene structure, expression, biochemical properties, and centromeric localization of its CENP-B. The amino acid sequence deduced from the cloned AGM CENP-B gene was established to be highly homologous to that of human and mouse CENP-B. In particular, the DNA binding and homodimer formation domains demonstrated 100{\%} identity to their human and mouse counterparts. Immunoblotting and DNA mobility shift analyses revealed CENP-B to be expressed in AGM cell lines. As predicted from the gene structure, the AGM CENP-B in the cell extracts exhibited the same DNA binding specificity and homodimer forming activity as human CENP-B. By indirect immunofluorescent staining of AGM mitotic cells with anti-CENP-B antibodies, a centromere-specific localization of AGM CENP- B could be demonstrated. We also isolated AGM α-satellite DNA with a CENP- B box-like sequence with CENP-B affinity. These results not only prove that CENP-B functionally persists in AGM cells but also suggest that the AGM genome contains the recognition sequences for CENP-B (CENP-B boxes with the core recognition sequence or CENP-B box variants) in centromeric satellite DNA.",
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