PT  - JOURNAL ARTICLE
AU  - Giuseppe Ciossani
AU  - Katharina Overlack
AU  - Arsen Petrovic
AU  - Pim Huis in ‘t Veld
AU  - Carolin Körner
AU  - Sabine Wohlgemuth
AU  - Stefano Maffini
AU  - Andrea Musacchio
TI  - Kinetochore recruitment of CENP-F illustrates how paralog divergence shapes kinetochore composition and function
AID  - 10.1101/276204
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 276204
4099  - http://biorxiv.org/content/early/2018/03/04/276204.short
4100  - http://biorxiv.org/content/early/2018/03/04/276204.full
AB  - The metazoan proteins CENP-E and CENP-F are components of a fibrous layer of mitotic kinetochores named the corona. Several features suggest that CENP-E and CENP-F are paralogs: they are very large (approximately 2700 and 3200 residues, respectively), rich in predicted coiled-coil structure, C-terminally prenylated, and endowed with microtubule-binding sites at their termini. In addition, CENP-E contains an ATP-hydrolyzing motor domain that promotes microtubule plus-end directed motion. Here, we show that CENP-E and CENP- F are recruited to mitotic kinetochores independently of the Rod-Zwilch-ZW10 (RZZ) complex, the main corona constituent. We identify selective interactions of CENP-E and CENP-F respectively with BubR1 and Bub1, paralogous proteins involved in mitotic checkpoint control and chromosome alignment. While BubR1 is dispensable for kinetochore localization of CENP-E, Bub1 is stringently required for CENP-F localization. Through biochemical reconstitution, we demonstrate that the CENP-E:BubR1 and CENP-F:Bub1 interactions are direct and require similar determinants, a dimeric coiled-coil in CENP-E or CENP-F and a kinase domain in BubR1 or Bub1. Our findings are consistent with the existence of ‘pseudo-symmetric’, paralogous Bub1:CENP-F and BubR1:CENP-E axes, supporting evolutionary relatedness of CENP-E and CENP-F.