RT Journal Article SR Electronic T1 Kinetochore recruitment of CENP-F illustrates how paralog divergence shapes kinetochore composition and function JF bioRxiv FD Cold Spring Harbor Laboratory SP 276204 DO 10.1101/276204 A1 Giuseppe Ciossani A1 Katharina Overlack A1 Arsen Petrovic A1 Pim Huis in ‘t Veld A1 Carolin Körner A1 Sabine Wohlgemuth A1 Stefano Maffini A1 Andrea Musacchio YR 2018 UL http://biorxiv.org/content/early/2018/03/04/276204.abstract 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.