Essentiality of CENP-A Depends on Its Binding Mode to HJURP
Tetsuya Hori,
JingHui Cao,
Kohei Nishimura,
Mariko Ariyoshi,
Yasuhiro Arimura,
Hitoshi Kurumizaka,
Tatsuo Fukagawa
Affiliations
Tetsuya Hori
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan; Corresponding author
JingHui Cao
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
Kohei Nishimura
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
Mariko Ariyoshi
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
Yasuhiro Arimura
Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
Hitoshi Kurumizaka
Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
Tatsuo Fukagawa
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan; Corresponding author
Summary: CENP-A incorporation is critical for centromere specification and is mediated by the chaperone HJURP. The CENP-A-targeting domain (CATD) of CENP-A specifically binds to HJURP, and this binding is conserved. However, the binding interface of CENP-A-HJURP is yet to be understood. Here, we identify the critical residues for chicken CENP-A or HJURP. The A59Q mutation in the α1-helix of chicken CENP-A causes CENP-A mis-incorporation and subsequent cell death, whereas the corresponding mutation in human CENP-A does not. We also find that W53 of HJURP, which is a contact site of A59 in CENP-A, is also essential in chicken cells. Our comprehensive analyses reveal that the affinities of HJURP to CATD differ between chickens and humans. However, the introduction of two arginine residues to the chicken HJURP αA-helix suppresses CENP-A mis-incorporation in chicken cells expressing CENP-AA59Q. Our data explain the mechanisms and evolution of CENP-A essentiality by the CENP-A-HJURP interaction.
