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bioRxiv · 10.1101/276204

Kinetochore recruitment of CENP-F illustrates how paralog divergence shapes kinetochore composition and function

Ciossani, G.·Overlack, K.·Petrovic, A.·Huis in 't Veld, P.·Koerner, C.·Wohlgemuth, S.·Maffini, S.·Musacchio, A.
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Abstract

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.

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Ciossani, G., Overlack, K., Petrovic, A., Huis in 't Veld, P., Koerner, C., Wohlgemuth, S., Maffini, S., Musacchio, A.. 2018-03-04. Kinetochore recruitment of CENP-F illustrates how paralog divergence shapes kinetochore composition and function. https://doi.org/10.1101/276204

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