Kinetochore-independent sister chromosome separation is driven by plus-end microtubule dynamics [article]

Hannah Vicars, Travis Karg, Brandt Warecki, Ian Bast, William Sullivan
2020 bioRxiv   pre-print
Although centromeres play a key role in sister chromatid separation and segregation, mitotic transmission of chromosome fragments lacking a centromere (acentrics) can be successful. In Drosophila neuroblasts, acentric chromosomes undergo delayed, but successful transmission revealing the existence of kinetochore-independent mechanisms driving transmission. In spite of delayed sister chromatid separation, bulk cohesin removal from the acentric is not delayed, suggesting factors other than
more » ... are responsible for the delay. In contrast to intact kinetochore-bearing chromosomes, we discovered that acentrics align parallel as well as perpendicular to the mitotic spindle. In addition, sister acentrics undergo unconventional patterns of separation. For example, rather than the simultaneous separation of sisters, acentrics oriented parallel to the spindle often slide past one another toward opposing poles. To identify the mechanisms driving acentric separation, we screened 117 RNAi gene knockdowns for synthetic lethality with acentric chromosome fragments. In addition to well-established DNA repair and checkpoint mutants, this candidate screen identified synthetic lethality with X-chromosome-derived acentric fragments in knockdowns of Greatwall (cell cycle kinase), EB1 (plus-end tracking protein), and Map205 (microtubule-stabilizing protein). Additional image-based screening revealed that reductions in Topoisomerase II levels disrupted sister acentric separation. Intriguingly, live imaging revealed that knockdowns of EB1, Map205, and Greatwall preferentially disrupted the sliding mode of sister acentric separation. In spite of these failures in acentric separation, acentrics incorporated into daughter nuclei and there was no increase in micronuclei formation. Based on our analysis of EB1 localization and knockdown phenotypes, we propose that in the absence of a kinetochore, microtubule plus-end dynamics provide the force to resolve DNA catenations required for sister separation.
doi:10.1101/2020.08.04.236943 fatcat:3thzi7a2svejhcs7aw5ds7a3xe