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CENP-C and CENP-I are key connecting factors for kinetochore and CENP-A assembly

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  • Nobuaki Shono
  • Jun Ichirou Ohzeki
  • Koichiro Otake
  • Nuno M C Martins
  • Takahiro Nagase
  • Hiroshi Kimura
  • Vladimir Larionov
  • William C. Earnshaw
  • Hiroshi Masumoto

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Original languageEnglish
Pages (from-to)4572-4587
Number of pages16
JournalJournal of Cell Science
Volume128
Issue number24
DOIs
Publication statusPublished - 2015

Abstract

Although it is generally accepted that chromatin containing the histone H3 variant CENP-A is an epigenetic mark maintaining centromere identity, the pathways leading to the formation and maintenance of centromere chromatin remain unclear.We previously generated human artificial chromosomes (HACs) whose centromeres contain a synthetic alpha-satellite (alphoid) DNA array containing the tetracycline operator (alphoidtetO).We also obtained cell lines bearing the alphoidtetO array at ectopic integration sites on chromosomal arms. Here, we have examined the regulation of CENP-A assembly at centromeres as well as de novo assembly on the ectopic arrays by tethering tetracycline repressor (tetR) fusions of substantial centromeric factors and chromatin modifiers. This analysis revealed four classes of factors that influence CENP-A assembly. Interestingly, many kinetochore structural components induced de novo CENP-A assembly at the ectopic site. We showed that these components work by recruiting CENP-C and subsequently recruiting M18BP1. Furthermore, we found that CENP-I can also recruit M18BP1 and, as a consequence, enhances M18BP1 assembly on centromeres in the downstream of CENP-C. Thus, we suggest that CENP-C and CENP-I are key factors connecting kinetochore to CENP-A assembly.

    Research areas

  • CENP-A, CENP-C, CENP-I, Centromere, HAC, Human artificial chromosome, Kinetochore

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