The Structural Basis for Kinetochore Stabilization by Cnn1/CENP-T

Hinshaw, Stephen M.

doi:10.1016/j.cub.2020.06.024

Cited by 21 publications

(29 citation statements)

References 61 publications

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“…To verify that the Ctf3-2A protein supports Ctf3c formation and kinetochore recruitment, we imaged cells expressing Mcm22-GFP (Figure 4A). Wild type cells showed Mcm22-GFP localization identical to what we have described for Ctf3-GFP (Hinshaw and Harrison, 2020). The signal in the ctf3-2A cells was indistinguishable from that observed in wild type cells, indicating that the Ctf3-2A protein does not disrupt Ctf3c assembly or kinetochore recruitment.…”

Section: Resultssupporting

confidence: 82%

“…A) Model of the assembled Ctf19c (Ctf3 – blue; Mcm22 – pink; Mcm16 – yellow; Cnn1/Wip – purple; Chl4, Iml3, Ctf19, Mcm21, Ame1, Okp1, Nkp1, Nkp2 – gray; EMD-0523, EMD-2910) (Hinshaw and Harrison, 2019, 2020). B) The Ctf3c binds the Ulp2-KIM but not the Ulp2-KIM-3A peptide.…”

Section: Resultsmentioning

confidence: 99%

“…D) Cartoon model showing Ulp2 association with the inner kinetochore. The image is adapted from Hinshaw and Harrison, 2020 (Cse4 nuc. – Cse4 nucleosome; CEN DNA – centromeric DNA).…”

Section: Resultsmentioning

confidence: 99%

“…Ctf3 mutations were created by two-step overlapping PCR amplification of the polycistronic insert before reinsertion into the original destination vector by ligationindependent cloning. Recombinant Cnn1-Wip1 protein sample used for cryo-EM was the same as used previously (Hinshaw and Harrison, 2020).…”

Section: Methodsmentioning

confidence: 99%

“…Inner kinetochore proteins, most of which assemble into the Ctf19 complex (Ctf19c in yeast, C onstitutive C entromere A ssociated N etwork or CCAN in vertebrates), contact centromeric DNA and regulate chromosomal functions. Structural studies of inner kinetochore proteins imply regulated chromatin recognition and show how key activities are coordinated (Hinshaw and Harrison, 2019, 2020; Kixmoeller et al, 2020; Yan et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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The structural basis for Ulp2 recruitment to the kinetochore

Quan

Hinshaw

Wang

et al. 2021

Preprint

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The step-by-step process of chromosome segregation defines the stages of the cell division cycle. In eukaryotes, signaling pathways that control these steps converge upon the kinetochore, a multiprotein assembly that connects spindle microtubules to the centromere of each chromosome. Kinetochores control and adapt to major chromosomal transactions, including replication of centromeric DNA, biorientation of sister centromeres on the metaphase spindle, and transit of sister chromatids into daughter cells during anaphase. Although the mechanisms that ensure tight microtubule coupling at anaphase are at least partly understood, kinetochore adaptations that support other cell cycle transitions are not. We report here a mechanism that enables regulated control of kinetochore sumoylation. A conserved surface of the Ctf3/CENP-I kinetochore protein provides a binding site for the SUMO protease, Ulp2. Ctf3 mutations that disable Ulp2 recruitment cause elevated inner kinetochore sumoylation and defective chromosome segregation. The location of the site within the assembled kinetochore suggests coordination between sumoylation and other cell cycle-regulated processes.

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Section: Resultssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

“…D) Cartoon model showing Ulp2 association with the inner kinetochore. The image is adapted from Hinshaw and Harrison, 2020 (Cse4 nuc. – Cse4 nucleosome; CEN DNA – centromeric DNA).…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The structural basis for Ulp2 recruitment to the kinetochore

Quan

Hinshaw

Wang

et al. 2021

Preprint

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Minichromosome maintenance proteins in eukaryotic chromosome segregation

et al. 2021

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Minichromosome maintenance (Mcm) proteins are well-known for their functions in DNA replication. However, their roles in chromosome segregation are yet to be reviewed in detail. Following the discovery in 1984, a group of Mcm proteins, known as the ARS-nonspecific group consisting of Mcm13, Mcm16-19, and Mcm21-22, were characterized as bonafide kinetochore proteins and were shown to play significant roles in the kinetochore assembly and high-fidelity chromosome segregation. This review focuses on the structure, function, and evolution of this group of Mcm proteins. Our in silico analysis of the physical interactors of these proteins reveals that they share non-overlapping functions despite being copurified in biochemically stable complexes. We have discussed the contrasting results reported in the literature and experimental strategies to address them. Taken together, this review focuses on the structure-function of the ARS-nonspecific Mcm proteins and their evolutionary flexibility to maintain genome stability in various organisms.

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