XB-ART-52049Chromosoma. March 1, 2017; 126 (2): 279-286.
Centromeric chromatin containing the histone H3 variant centromere protein A (CENP-A) directs kinetochore assembly through a hierarchical binding of CENPs, starting with CENP-C and CENP-T. Centromeres are also the chromosomal regions where cohesion, mediated by cohesin, is most prominently maintained in mitosis. While most cohesin dissociates from chromosome arms in prophase, Shugoshin 1 (Sgo1) prevents this process at centromeres. Centromeric localization of Sgo1 depends on histone H2A phosphorylation by the kinase Bub1, but whether additional interactions with kinetochore components are required for Sgo1 recruitment is unclear. Using the Xenopus egg cell-free system, we here show that both CENP-C and CENP-T can independently drive centromeric accumulation of Sgo1 through recruitment of Bub1 to the KNL1, MIS12, NDC80 (KMN) network. The spindle assembly checkpoint (SAC) kinase Mps1 is also required for this pathway even in the absence of checkpoint signaling. Sgo1 recruitment is abolished in chromosomes lacking kinetochore components other than CENP-A. However, forced targeting of Bub1 to centromeres is sufficient to restore Sgo1 localization under this condition.
PubMed ID: 27116032
PMC ID: PMC5371614
Article link: Chromosoma.
Genes referenced: bub1 cenpa cenpc cenpt kidins220 rbbp4 rps27 sgo1
Article Images: [+] show captions
|Fig. 2. Reduced Sgo1 recruitment to chromatin in the absence of CENP-C, CENP-T, Bub1, or Mps1. Immunoblot analysis of chromatin fractions from replicated chromosomes assembled in the indicated extracts and purified by centrifugation through a sucrose cushion (lanes 2–7). Chromatin purified in the same way from a mock assembly reaction without sperm served as control (no sp, lane 1). Histone H1 was used as loading control. Quantification of the Sgo1 signals, normalized to the H1 signals, and expressed relative to the Sgo1 signal in the chromatin obtained in the mock-depleted extract|