PT  - JOURNAL ARTICLE
AU  - Honghui Jiang
AU  - Mariko Ariyoshi
AU  - Reito Watanabe
AU  - Fumiaki Makino
AU  - Keiichi Namba
AU  - Tatsuo Fukagawa
TI  - The cryo-EM structure of the CENP-A nucleosome in complex with ggKNL2
AID  - 10.1101/2022.06.24.497480
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.06.24.497480
4099  - http://biorxiv.org/content/early/2022/06/24/2022.06.24.497480.short
4100  - http://biorxiv.org/content/early/2022/06/24/2022.06.24.497480.full
AB  - Centromere protein A (CENP-A) nucleosome is an epigenetic marker that specifies centromere position. The Mis18 complex is a licensing factor for new CENP-A deposition via the CENP-A chaperone, Holliday junction recognition protein (HJURP) on the centromere chromatin. Chicken KINETOCHORE NULL2 (KNL2) (ggKNL2), a Mis18 complex component, has a CENP-C-like motif, and our previous study suggested that ggKNL2 directly binds to the CENP-A nucleosome to recruit HJURP/CENP-A to the centromere. However, the molecular basis for CENP-A nucleosome recognition by ggKNK2 remains unclear. Here, we present the cryo-EM structure of the chicken CENP-A nucleosome in complex with a ggKNL2 fragment containing a CENP-C-like motif. Chicken KNL2 distinguishes between CENP-A and histone H3 in the nucleosome using the CENP-C-like motif and its downstream region. Both the C-terminal tail and RG-loop of CENP-A are simultaneously recognized as CENP-A characteristics. The CENP-A nucleosome-ggKNL2 interaction is thus essential for CENP-A deposition. Furthermore, our structural, biochemical, and cell biology data indicate that ggKNL2 alters its binding partner at the centromere during chicken cell cycle progression.