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  1. Article: Modification of the Neck Linker of KIF18A Alters Microtubule Subpopulation Preference.

    Queen, Katelyn A / Cario, Alisa / Berger, Christopher L / Stumpff, Jason

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Kinesins support many diverse cellular processes, including facilitating cell division through mechanical regulation of the mitotic spindle. However, how kinesin activity is controlled to facilitate this process is not well understood. Interestingly, ... ...

    Abstract Kinesins support many diverse cellular processes, including facilitating cell division through mechanical regulation of the mitotic spindle. However, how kinesin activity is controlled to facilitate this process is not well understood. Interestingly, post-translational modifications have been identified within the enzymatic region of all 45 mammalian kinesins, but the significance of these modifications has gone largely unexplored. Given the critical role of the enzymatic region in facilitating nucleotide and microtubule binding, it may serve as a primary site for kinesin regulation. Consistent with this idea, a phosphomimetic mutation at S357 in the neck-linker of KIF18A alters the localization of KIF18A within the spindle from kinetochore microtubules to peripheral microtubules. Changes in localization of KIF18A-S357D are accompanied by defects in mitotic spindle positioning and the ability to promote mitotic progression. This altered localization pattern is mimicked by a shortened neck-linker mutant, suggesting that KIF18A-S357D may cause the motor to adopt a shortened neck-linker like state that prevents KIF18A from accumulating at the plus-ends of kinetochore microtubules. These findings demonstrate that post-translational modifications in the enzymatic region of kinesins could be important for biasing their localization to particular microtubule subpopulations.
    Language English
    Publishing date 2023-05-02
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.05.02.539080
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Modification of the neck-linker of KIF18A alters Microtubule subpopulation preference.

    Queen, Katelyn A / Cario, Alisa / Berger, Christopher L / Stumpff, Jason

    Molecular biology of the cell

    2023  Volume 35, Issue 1, Page(s) ar3

    Abstract: Kinesins support many diverse cellular processes, including facilitating cell division through mechanical regulation of the mitotic spindle. However, how kinesin activity is controlled to facilitate this process is not well understood. Interestingly, ... ...

    Abstract Kinesins support many diverse cellular processes, including facilitating cell division through mechanical regulation of the mitotic spindle. However, how kinesin activity is controlled to facilitate this process is not well understood. Interestingly, posttranslational modifications have been identified within the enzymatic region of all 45 mammalian kinesins, but the significance of these modifications has gone largely unexplored. Given the critical role of the enzymatic region in facilitating nucleotide and microtubule binding, it may serve as a primary site for kinesin regulation. Consistent with this idea, a phosphomimetic mutation at S357 in the neck-linker of KIF18A alters the localization of KIF18A within the spindle from kinetochore microtubules to nonkinetochore microtubules at the periphery of the spindle. Changes in localization of KIF18A-S357D are accompanied by defects in mitotic spindle positioning and the ability to promote mitotic progression. This altered localization pattern is mimicked by a shortened neck-linker mutant, suggesting that KIF18A-S357D may cause the motor to adopt a shortened neck-linker-like state that decreases KIF18A accumulation at the plus-ends of kinetochore microtubules. These findings demonstrate that posttranslational modifications in the enzymatic region of kinesins could be important for biasing their localization to particular microtubule subpopulations.
    MeSH term(s) Animals ; Humans ; HeLa Cells ; Kinesins/metabolism ; Kinetochores/metabolism ; Microtubules/metabolism ; Mitosis ; Spindle Apparatus/metabolism
    Chemical Substances KIF18A protein, human (EC 3.6.1.-) ; Kinesins (EC 3.6.4.4)
    Language English
    Publishing date 2023-10-30
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1098979-1
    ISSN 1939-4586 ; 1059-1524
    ISSN (online) 1939-4586
    ISSN 1059-1524
    DOI 10.1091/mbc.E23-05-0167
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Quantifying Changes in Chromosome Position to Assess Chromokinesin Activity.

    Thompson, Alex F / Vandal, Sarah / Stumpff, Jason

    Methods in molecular biology (Clifton, N.J.)

    2021  Volume 2415, Page(s) 139–149

    Abstract: The chromokinesin KIF22 (Kid, kinesin-10 family) is the primary generator of polar ejection forces, which contribute to chromosome positioning and alignment in mitotic cells. Assessment of KIF22 function requires quantitative comparison of relative polar ...

    Abstract The chromokinesin KIF22 (Kid, kinesin-10 family) is the primary generator of polar ejection forces, which contribute to chromosome positioning and alignment in mitotic cells. Assessment of KIF22 function requires quantitative comparison of relative polar ejection forces between experimental conditions. This is facilitated by the generation of monopolar spindles to reduce the impact of bioriented microtubule attachment at kinetochores on chromosome positions and increase the dependence of chromosome positions on chromokinesin activity. Radial profile plots measure the intensity of chromatin signal in concentric circles around the poles of monopolar cells and represent an expedient quantitative measure of relative polar ejection forces. As such, this assay can be used to measure changes in polar ejection forces resulting from chromokinesin depletion or perturbation.
    MeSH term(s) Chromosomes/genetics ; DNA-Binding Proteins/genetics ; Kinesins ; Kinetochores ; Microtubules ; Mitosis ; Nuclear Proteins/genetics ; Spindle Apparatus
    Chemical Substances DNA-Binding Proteins ; Nuclear Proteins ; chromokinesin ; Kinesins (EC 3.6.4.4)
    Language English
    Publishing date 2021-12-07
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-1904-9_10
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Measuring microtubule thickness: an exercise in cooperativity.

    Stumpff, Jason

    Developmental cell

    2012  Volume 23, Issue 1, Page(s) 1–2

    Abstract: Doublecortin (DCX), a microtubule-associated protein, is essential for neuronal migration, although a clear mechanistic understanding of this requirement remains elusive. In this issue of Developmental Cell, Bechstedt and Brouhard (2012) report that DCX ... ...

    Abstract Doublecortin (DCX), a microtubule-associated protein, is essential for neuronal migration, although a clear mechanistic understanding of this requirement remains elusive. In this issue of Developmental Cell, Bechstedt and Brouhard (2012) report that DCX relies on cooperative binding and an affinity for growing microtubule ends to nucleate and stabilize 13-protofilament microtubules.
    Language English
    Publishing date 2012-07-17
    Publishing country United States
    Document type Comment ; Journal Article
    ZDB-ID 2054967-2
    ISSN 1878-1551 ; 1534-5807
    ISSN (online) 1878-1551
    ISSN 1534-5807
    DOI 10.1016/j.devcel.2012.06.010
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Identification of the KIF18A alpha-4 helix as a therapeutic target for chromosomally unstable tumor cells.

    Schutt, Katherine L / Queen, Katelyn A / Fisher, Kira / Budington, Olivia / Mao, Weifeng / Liu, Wei / Gu, Xiaohui / Xiao, Yisong / Aswad, Fred / Joseph, James / Stumpff, Jason

    Frontiers in molecular biosciences

    2024  Volume 11, Page(s) 1328077

    Abstract: Background: ...

    Abstract Background:
    Language English
    Publishing date 2024-02-12
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2814330-9
    ISSN 2296-889X
    ISSN 2296-889X
    DOI 10.3389/fmolb.2024.1328077
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Measuring Microtubule Thickness: An Exercise in Cooperativity

    Stumpff, Jason

    Developmental cell. 2012 July 17, v. 23, no. 1

    2012  

    Abstract: Doublecortin (DCX), a microtubule-associated protein, is essential for neuronal migration, although a clear mechanistic understanding of this requirement remains elusive. In this issue of Developmental Cell, Bechstedt and Brouhard (2012) report that DCX ... ...

    Abstract Doublecortin (DCX), a microtubule-associated protein, is essential for neuronal migration, although a clear mechanistic understanding of this requirement remains elusive. In this issue of Developmental Cell, Bechstedt and Brouhard (2012) report that DCX relies on cooperative binding and an affinity for growing microtubule ends to nucleate and stabilize 13-protofilament microtubules.
    Keywords binding capacity ; cell movement ; microtubules ; neurons ; proteins
    Language English
    Dates of publication 2012-0717
    Size p. 1-2.
    Publishing place Elsevier Inc.
    Document type Article
    ZDB-ID 2054967-2
    ISSN 1878-1551 ; 1534-5807
    ISSN (online) 1878-1551
    ISSN 1534-5807
    DOI 10.1016/j.devcel.2012.06.010
    Database NAL-Catalogue (AGRICOLA)

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  7. Article ; Online: Quantification of Mitotic Chromosome Alignment.

    Fonseca, Cindy / Stumpff, Jason

    Methods in molecular biology (Clifton, N.J.)

    2016  Volume 1413, Page(s) 253–262

    Abstract: The alignment of chromosomes during metaphase is a hallmark of mitosis. For this reason, chromosome alignment has served as an informative functional assay for evaluating mitotic fidelity. The common approach of quantifying the number of mitotic cells ... ...

    Abstract The alignment of chromosomes during metaphase is a hallmark of mitosis. For this reason, chromosome alignment has served as an informative functional assay for evaluating mitotic fidelity. The common approach of quantifying the number of mitotic cells with unaligned chromosomes within a population has led to the identification of many proteins required for this conserved process. However, more sensitive assays are now required to dissect the complex molecular control of chromosome alignment. In this chapter, we describe a microscopy-based method for objectively quantifying the distribution of fluorescently labeled chromosomes within the mitotic spindle that can be used to evaluate the extent of chromosome alignment within individual mitotic cells.
    Language English
    Publishing date 2016
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-3542-0_16
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Micronuclei in Kif18a mutant mice form stable micronuclear envelopes and do not promote tumorigenesis.

    Sepaniac, Leslie A / Martin, Whitney / Dionne, Louise A / Stearns, Timothy M / Reinholdt, Laura G / Stumpff, Jason

    The Journal of cell biology

    2021  Volume 220, Issue 11

    Abstract: Micronuclei, whole or fragmented chromosomes spatially separated from the main nucleus, are associated with genomic instability and have been identified as drivers of tumorigenesis. Paradoxically, Kif18a mutant mice produce micronuclei due to ... ...

    Abstract Micronuclei, whole or fragmented chromosomes spatially separated from the main nucleus, are associated with genomic instability and have been identified as drivers of tumorigenesis. Paradoxically, Kif18a mutant mice produce micronuclei due to asynchronous segregation of unaligned chromosomes in vivo but do not develop spontaneous tumors. We report here that micronuclei in Kif18a mutant mice form stable nuclear envelopes. Challenging Kif18a mutant mice via deletion of the Trp53 gene led to formation of thymic lymphoma with elevated levels of micronuclei. However, loss of Kif18a had modest or no effect on survival of Trp53 homozygotes and heterozygotes, respectively. Micronuclei in cultured KIF18A KO cells form stable nuclear envelopes characterized by increased recruitment of nuclear envelope components and successful expansion of decondensing chromatin compared with those induced by nocodazole washout or radiation. Lagging chromosomes were also positioned closer to the main chromatin masses in KIF18A KO cells. These data suggest that not all micronuclei actively promote tumorigenesis.
    MeSH term(s) Animals ; Carcinogenesis/genetics ; Cell Line ; Cell Nucleus/genetics ; Chromatin/genetics ; Chromosomes/genetics ; DNA Damage/genetics ; Female ; Genomic Instability/genetics ; Humans ; Kinesins/genetics ; Male ; Mice ; Nuclear Envelope/genetics
    Chemical Substances Chromatin ; KIF18a protein, mouse ; Kinesins (EC 3.6.4.4)
    Language English
    Publishing date 2021-09-13
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.202101165
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Pathogenic mutations in the chromokinesin KIF22 disrupt anaphase chromosome segregation.

    Thompson, Alex F / Blackburn, Patrick R / Arons, Noah S / Stevens, Sarah N / Babovic-Vuksanovic, Dusica / Lian, Jane B / Klee, Eric W / Stumpff, Jason

    eLife

    2022  Volume 11

    Abstract: The chromokinesin KIF22 generates forces that contribute to mitotic chromosome congression and alignment. Mutations in the α2 helix of the motor domain of KIF22 have been identified in patients with abnormal skeletal development, and we report the ... ...

    Abstract The chromokinesin KIF22 generates forces that contribute to mitotic chromosome congression and alignment. Mutations in the α2 helix of the motor domain of KIF22 have been identified in patients with abnormal skeletal development, and we report the identification of a patient with a novel mutation in the KIF22 tail. We demonstrate that pathogenic mutations do not result in a loss of KIF22's functions in early mitosis. Instead, mutations disrupt chromosome segregation in anaphase, resulting in reduced proliferation, abnormal daughter cell nuclear morphology, and, in a subset of cells, cytokinesis failure. This phenotype could be explained by a failure of KIF22 to inactivate in anaphase. Consistent with this model, constitutive activation of the motor via a known site of phosphoregulation in the tail phenocopied the effects of pathogenic mutations. These results suggest that the motor domain α2 helix may be an important site for regulation of KIF22 activity at the metaphase to anaphase transition. In support of this conclusion, mimicking phosphorylation of α2 helix residue T158 also prevents inactivation of KIF22 in anaphase. These findings demonstrate the importance of both the head and tail of the motor in regulating the activity of KIF22 and offer insight into the cellular consequences of preventing KIF22 inactivation and disrupting force balance in anaphase.
    MeSH term(s) Anaphase ; Chromosome Segregation ; DNA-Binding Proteins/genetics ; Kinesins/genetics ; Metaphase ; Mitosis ; Mutation ; Nuclear Proteins/genetics ; Spindle Apparatus
    Chemical Substances DNA-Binding Proteins ; Nuclear Proteins ; chromokinesin ; Kinesins (EC 3.6.4.4)
    Language English
    Publishing date 2022-06-22
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.78653
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: KIF18A's neck linker permits navigation of microtubule-bound obstacles within the mitotic spindle.

    Malaby, Heidi Lh / Lessard, Dominique V / Berger, Christopher L / Stumpff, Jason

    Life science alliance

    2019  Volume 2, Issue 1

    Abstract: KIF18A (kinesin-8) is required for mammalian mitotic chromosome alignment. KIF18A confines chromosome movement to the mitotic spindle equator by accumulating at the plus-ends of kinetochore microtubule bundles (K-fibers), where it functions to suppress K- ...

    Abstract KIF18A (kinesin-8) is required for mammalian mitotic chromosome alignment. KIF18A confines chromosome movement to the mitotic spindle equator by accumulating at the plus-ends of kinetochore microtubule bundles (K-fibers), where it functions to suppress K-fiber dynamics. It is not understood how the motor accumulates at K-fiber plus-ends, a difficult feat requiring the motor to navigate protein dense microtubule tracks. Our data indicate that KIF18A's relatively long neck linker is required for the motor's accumulation at K-fiber plus-ends. Shorter neck linker (sNL) variants of KIF18A display a deficiency in accumulation at the ends of K-fibers at the center of the spindle. Depletion of K-fiber-binding proteins reduces the KIF18A sNL localization defect, whereas their overexpression reduces wild-type KIF18A's ability to accumulate on this same K-fiber subset. Furthermore, single-molecule assays indicate that KIF18A sNL motors are less proficient in navigating microtubules coated with microtubule-associated proteins. Taken together, these results support a model in which KIF18A's neck linker length permits efficient navigation of obstacles to reach K-fiber ends during mitosis.
    MeSH term(s) Chromosomes/metabolism ; HeLa Cells ; Humans ; Kinesin/genetics ; Kinesin/metabolism ; Kinetochores/metabolism ; Leupeptins/pharmacology ; Metaphase/drug effects ; Metaphase/physiology ; Microtubules/metabolism ; RNA, Small Interfering/genetics ; Spindle Apparatus/metabolism ; Transfection
    Chemical Substances Leupeptins ; RNA, Small Interfering ; KIF18A protein, human (EC 3.6.1.-) ; Kinesin (EC 3.6.4.4) ; benzyloxycarbonylleucyl-leucyl-leucine aldehyde (RF1P63GW3K)
    Language English
    Publishing date 2019-01-17
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ISSN 2575-1077
    ISSN (online) 2575-1077
    DOI 10.26508/lsa.201800169
    Database MEDical Literature Analysis and Retrieval System OnLINE

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