Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells

Cao, Jing Hui; Hori, Tetsuya; Ariyoshi, Mariko; Fukagawa, Tatsuo

doi:https://doi.org/10.1242/jcs.261639

Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells

Cao, Jing Hui; Hori, Tetsuya; Ariyoshi, Mariko et al.

Journal of Cell Science, 2024, 137(3)

アクセス数:70件（2025-05-19 14:23 集計）

固定URL: https://hdl.handle.net/11094/96426

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論文情報

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タイトル	Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells
著者	著者名 Cao, Jing Hui
	NRID 1000070550078 1000070550078 researchmap permalink thori567 thori567 Scopus Author ID 7402411497 7402411497 著者名 Hori, Tetsuya
	NRID 1000080437243 1000080437243 ORCID 0000-0002-1361-2642 0000-0002-1361-2642 researchmap permalink 7000010594 7000010594 Scopus Author ID 7006547610 7006547610 著者名 Ariyoshi, Mariko
	NRID 1000060321600 1000060321600 researchmap permalink fukagawa_tatsuo fukagawa_tatsuo Scopus Author ID 7003912496 7003912496 著者名 Fukagawa, Tatsuo
キーワード等	CENP-A
	CENP-C
	Centromere
	HJURP
	Kinetochore
	Knl2
抄録	The kinetochore is an essential structure for chromosome segregation. Although the kinetochore is usually formed on a centromere locus, it can be artificially formed at a non-centromere locus by protein tethering. An artificial kinetochore can be formed by tethering of CENP-C or CENP-I, members of the constitutive centromere-associated network (CCAN). However, how CENP-C or CENP-I recruit the centromere-specific histone CENP-A to form an artificial kinetochore remains unclear. In this study, we analyzed this issue using the tethering assay combined with an auxin-inducible degron (AID)-based knockout method in chicken DT40 cells. We found that tethering of CENP-C or CENP-I induced CENP-A incorporation at the non-centromeric locus in the absence of Knl2 (or MIS18BP1), a component of the Mis18 complex, and that Knl2 tethering recruited CENP-A in the absence of CENP-C. We also showed that CENP-C coimmunoprecipitated with HJURP, independently of Knl2. Considering these results, we propose that CENP-C recruits CENP-A by HJURP binding to form an artificial kinetochore. Our results suggest that CENP-C or CENP-I exert CENP-A recruitment activity, independently of Knl2, for artificial kinetochore formation in chicken DT40 cells. This gives us a new insight into mechanisms for CENP-A incorporation.
出版者	Company of Biologists Ltd
収録物名	Journal of Cell Science
巻(号)	137(3)
刊行年月	2024-02-06
言語	英語
ハンドルURL	https://hdl.handle.net/11094/96426
PISSN	00219533
eISSN	14779137
関連情報 (isVersionOf)	DOI (出版社版) https://doi.org/10.1242/jcs.261639
アクセス権	open access
出版タイプ	Accepted Manuscript
カテゴリ	学術雑誌論文 Journal Article

論文詳細を表示

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCTERMS.bibliographicCitation	Journal of Cell Science. 2024, 137(3)
DC.title	Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells
DC.creator	Cao, Jing Hui
	Hori, Tetsuya
	Ariyoshi, Mariko
	Fukagawa, Tatsuo
DC.publisher	Company of Biologists Ltd
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2024-02-06
DC.identifier	https://doi.org/10.1242/jcs.261639
DC.subject	CENP-A
	CENP-C
	Centromere
	HJURP
	Kinetochore
	Knl2
DCTERMS.abstract	The kinetochore is an essential structure for chromosome segregation. Although the kinetochore is usually formed on a centromere locus, it can be artificially formed at a non-centromere locus by protein tethering. An artificial kinetochore can be formed by tethering of CENP-C or CENP-I, members of the constitutive centromere-associated network (CCAN). However, how CENP-C or CENP-I recruit the centromere-specific histone CENP-A to form an artificial kinetochore remains unclear. In this study, we analyzed this issue using the tethering assay combined with an auxin-inducible degron (AID)-based knockout method in chicken DT40 cells. We found that tethering of CENP-C or CENP-I induced CENP-A incorporation at the non-centromeric locus in the absence of Knl2 (or MIS18BP1), a component of the Mis18 complex, and that Knl2 tethering recruited CENP-A in the absence of CENP-C. We also showed that CENP-C coimmunoprecipitated with HJURP, independently of Knl2. Considering these results, we propose that CENP-C recruits CENP-A by HJURP binding to form an artificial kinetochore. Our results suggest that CENP-C or CENP-I exert CENP-A recruitment activity, independently of Knl2, for artificial kinetochore formation in chicken DT40 cells. This gives us a new insight into mechanisms for CENP-A incorporation.
DC.format	96426
citation_title	Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells
citation_author	Cao, Jing Hui
	Hori, Tetsuya
	Ariyoshi, Mariko
	Fukagawa, Tatsuo
citation_publisher	Company of Biologists Ltd
citation_language	英語
citation_date	2024-02-06
citation_journal_title	Journal of Cell Science
citation_volume	137
citation_issue	3
citation_doi	https://doi.org/10.1242/jcs.261639
citation_public_url	https://hdl.handle.net/11094/96426