CENP-F stabilizes kinetochore-microtubule attachments and limits dynein stripping of corona cargoes

Abstract

Accurate chromosome segregation demands efficient capture of microtubules by kinetochores and their conversion to stable bioriented attachments that can congress and then segregate chromosomes. An early event is the shedding of the outermost fibrous corona layer of the kinetochore following microtubule attachment. Centromere protein F (CENP-F) is part of the corona, contains two microtubule-binding domains, and physically associates with dynein motor regulators. Here, we have combined CRISPR gene editing and engineered separation-of-function mutants to define how CENP-F contributes to kinetochore function. We show that the two microtubule-binding domains make distinct contributions to attachment stability and force transduction but are dispensable for chromosome congression. We further identify a specialized domain that functions to limit the dynein-mediated stripping of corona cargoes through a direct interaction with Nde1. This antagonistic activity is crucial for maintaining the required corona composition and ensuring efficient kinetochore biorientation.

Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores

“…This conversion engages the core microtubule receptor of the kinetochore, the NDC80 complex, a sub-complex of the KMN network. A crucial aspect of the lateral to end-on conversion is that it coincides with a sudden activation of DD and with the disassembly of the kinetochore corona, in a process traditionally known as “shedding” (Auckland et al, 2020; Basto et al, 2004; Howell et al, 2001; Mische et al, 2008; Sivaram et al, 2009; Varma et al, 2008; Williams et al, 1996; Wojcik et al, 2001).…”

The Role of Mitotic Kinases and the RZZ Complex in Kinetochore-Microtubule Attachments: Doing the Right Link

“…Curiously, preventing CENP-F:NDE1 from blocking excessive removal of corona components, including the RZZc, caused an increase in chromosome segregation errors in human HeLa cells. Despite the rapid loss of the RZZc-and the fibrous corona -, these cells had only milder defects in chromosome congression, arguing that chromosome capture was not severely impaired (Auckland et al, 2020). Therefore, the observed increased rate of chromosome mis-segregation could potentially result from a deficient RZZc-mediated regulation of KT-MT attachment fidelity.…”

“…CENP-F has also been suggested to restrict DYNEIN-mediated removal of corona components in a NDE1dependent manner. However, a CENP-F:NDE1-dependent mechanism appears to regulate the release of several DYNEIN cargos, indicating that it broadly regulates DYNEIN function in fibrous corona assembly dynamics (Auckland et al, 2020). Curiously, preventing CENP-F:NDE1 from blocking excessive removal of corona components, including the RZZc, caused an increase in chromosome segregation errors in human HeLa cells.…”

Anti-centromere antibodies target centromere–kinetochore macrocomplex: a comprehensive autoantigen profiling

“…Autoimmunity centromere-kinetochore-microtubule interface is described in figure 1 based on the cited references. [6][7][8][9][10][11] Among a number of component molecules, CENP-A, CENP-B, CENP-C and CBX5 are known targets of ACAs. 12 13 In particular, CENP-B is thought to be the main autoantigen because the presence of anti-CENP-B antibody is highly consistent with the ACAs detected by the ANA test.…”