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  • The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore.

The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore.

eLife (2019-05-22)
Josef Fischböck-Halwachs, Sylvia Singh, Mia Potocnjak, Götz Hagemann, Victor Solis-Mezarino, Stephan Woike, Medini Ghodgaonkar-Steger, Florian Weissmann, Laura D Gallego, Julie Rojas, Jessica Andreani, Alwin Köhler, Franz Herzog
ABSTRACT

Kinetochores are macromolecular protein complexes at centromeres that ensure accurate chromosome segregation by attaching chromosomes to spindle microtubules and integrating safeguard mechanisms. The inner kinetochore is assembled on CENP-A nucleosomes and has been implicated in establishing a kinetochore-associated pool of Aurora B kinase, a chromosomal passenger complex (CPC) subunit, which is essential for chromosome biorientation. By performing crosslink-guided in vitro reconstitution of budding yeast kinetochore complexes we showed that the Ame1/Okp1CENP-U/Q heterodimer, which forms the COMA complex with Ctf19/Mcm21CENP-P/O, selectively bound Cse4CENP-A nucleosomes through the Cse4 N-terminus. The Sli15/Ipl1INCENP/Aurora-B core-CPC interacted with COMA in vitro through the Ctf19 C-terminus whose deletion affected chromosome segregation fidelity in Sli15 wild-type cells. Tethering Sli15 to Ame1/Okp1 rescued synthetic lethality upon Ctf19 depletion in a Sli15 centromere-targeting deficient mutant. This study shows molecular characteristics of the point-centromere kinetochore architecture and suggests a role for the Ctf19 C-terminus in mediating CPC-binding and accurate chromosome segregation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Millipore
ANTI-FLAG® M1 Agarose Affinity Gel