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)

Number of Access:427（2025-07-27 21:14 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/96426

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Title
Artificial tethering of constitutive centromere-associated network proteins induces CENP-A deposition without Knl2 in DT40 cells
Creator
CreatorName
Cao, Jing Hui
NRID
1000070550078 1000070550078
researchmap permalink
thori567 thori567
Scopus Author ID
7402411497 7402411497
CreatorName
Hori, Tetsuya
NRID
1000080437243 1000080437243
ORCID
0000-0002-1361-2642 0000-0002-1361-2642
researchmap permalink
7000010594 7000010594
Scopus Author ID
7006547610 7006547610
CreatorName
Ariyoshi, Mariko
NRID
1000060321600 1000060321600
researchmap permalink
fukagawa_tatsuo fukagawa_tatsuo
Scopus Author ID
7003912496 7003912496
CreatorName
Fukagawa, Tatsuo
Subject
CENP-A
CENP-C
Centromere
HJURP
Kinetochore
Knl2
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.
Publisher
Company of Biologists Ltd
Source Title
Journal of Cell Science
Volume (Issue)
137(3)
Date of Issued
2024-02-06
Language
English
Handle URL
https://hdl.handle.net/11094/96426
PISSN
00219533
eISSN
14779137
Relation.isVersionOf
DOI (Publisher Version)
https://doi.org/10.1242/jcs.261639
Access Rights
open access
oaire:version
Accepted Manuscript
Category
学術雑誌論文 Journal Article