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TitleElectron cryotomography analysis of Dam1C/DASH at the kinetochore-spindle interface in situ.
Journal, issue, pagesJ Cell Biol, Vol. 218, Issue 2, Page 455-473, Year 2019
Publish dateFeb 4, 2019
AuthorsCai Tong Ng / Li Deng / Chen Chen / Hong Hwa Lim / Jian Shi / Uttam Surana / Lu Gan /
PubMed AbstractIn dividing cells, depolymerizing spindle microtubules move chromosomes by pulling at their kinetochores. While kinetochore subcomplexes have been studied extensively in vitro, little is known about ...In dividing cells, depolymerizing spindle microtubules move chromosomes by pulling at their kinetochores. While kinetochore subcomplexes have been studied extensively in vitro, little is known about their in vivo structure and interactions with microtubules or their response to spindle damage. Here we combine electron cryotomography of serial cryosections with genetic and pharmacological perturbation to study the yeast chromosome segregation machinery in vivo. Each kinetochore microtubule has one (rarely, two) Dam1C/DASH outer kinetochore assemblies. Dam1C/DASH contacts the microtubule walls and does so with its flexible "bridges"; there are no contacts with the protofilaments' curved tips. In metaphase, ∼40% of the Dam1C/DASH assemblies are complete rings; the rest are partial rings. Ring completeness and binding position along the microtubule are sensitive to kinetochore attachment and tension, respectively. Our study and those of others support a model in which each kinetochore must undergo cycles of conformational change to couple microtubule depolymerization to chromosome movement.
External linksJ Cell Biol / PubMed:30504246 / PubMed Central
MethodsEM (tomography)
Resolution32.0 Å
Structure data

EMDB-6912:
Subtomogram average of the budding yeast DASH outer-kinetochore ring.
Method: EM (tomography) / Resolution: 32.0 Å

EMDB-6914:
Serial cryotomogram of a metaphase budding yeast cell.
Method: EM (tomography)

Source
  • Escherichia coli BL21(DE3) (bacteria)
  • Saccharomyces cerevisiae (brewer's yeast)

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