TY - JOUR
T1 - The COMA complex interacts with Cse4 and positions Sli15/ipl1 at the budding yeast inner kinetochore
AU - Fischböck-Halwachs, Josef
AU - Singh, Sylvia
AU - Potocnjak, Mia
AU - Hagemann, Götz
AU - Solis-Mezarino, Victor
AU - Woike, Stephan
AU - Ghodgaonkar-Steger, Medini
AU - Weissmann, Florian
AU - Gallego, Laura D.
AU - Rojas, Julie
AU - Andreani, Jessica
AU - Köhler, Alwin
AU - Herzog, Franz
N1 - Funding Information:
We are grateful to Andrea Musacchio (MPI Dortmund) and Stefan Westermann (University of Essen) for discussions and sharing reagents. We thank Wolfgang Zachariae (MPI Munich) for help with fluorescence microscopy. JFH and GH were funded by the Graduate School (GRK 1721) and MP and VS were funded by the Graduate School (Quantitative Biosciences Munich) of the German Research Foundation (DFG). LDG was a recipient of a DOC Fellowship of the Austrian Academy of Sciences and AK was funded by ERC Grant 281354 (NPC GENEXPRESS). FH was supported by the European Research Council (ERC-StG no. 638218), the Human Frontier Science Program (RGP0008/2015), by the Bavarian Research Center of Molecular Biosystems and by an LMU excellent junior grant.
Publisher Copyright:
© Fischböck-Halwachs et al.
PY - 2019/5/21
Y1 - 2019/5/21
N2 - 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.
AB - 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.
KW - Kinetochores/chemistry
KW - Protein Binding
KW - Protein Interaction Maps
KW - Saccharomyces cerevisiae Proteins/analysis
KW - Saccharomycetales/chemistry
UR - http://www.scopus.com/inward/record.url?scp=85067086777&partnerID=8YFLogxK
U2 - 10.7554/eLife.42879
DO - 10.7554/eLife.42879
M3 - Article
C2 - 31112132
AN - SCOPUS:85067086777
SN - 2050-084X
VL - 8
JO - eLife
JF - eLife
M1 - e42879
ER -