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  1. Article ; Online: A celebration of the 25th anniversary of chromatin-mediated spindle assembly.

    Verma, Vikash / Maresca, Thomas J

    Molecular biology of the cell

    2022  Volume 33, Issue 2, Page(s) rt1

    Abstract: Formation of a bipolar spindle is required for the faithful segregation of chromosomes during cell division. Twenty-five years ago, a transformative insight into how bipolarity is achieved was provided by Rebecca Heald, Eric Karsenti, and colleagues in ... ...

    Abstract Formation of a bipolar spindle is required for the faithful segregation of chromosomes during cell division. Twenty-five years ago, a transformative insight into how bipolarity is achieved was provided by Rebecca Heald, Eric Karsenti, and colleagues in their landmark publication characterizing a chromatin-mediated spindle assembly pathway in which centrosomes and kinetochores were dispensable. The discovery revealed that bipolar spindle assembly is a self-organizing process where microtubules, which possess an intrinsic polarity, polymerize around chromatin and become sorted by mitotic motors into a bipolar structure. On the 25
    MeSH term(s) Animals ; Anniversaries and Special Events ; Cell Cycle ; Centrosome ; Chromatin/metabolism ; Chromatin Assembly and Disassembly/physiology ; Humans ; Kinetochores ; Microtubules/metabolism ; Mitosis ; Spindle Apparatus/physiology
    Chemical Substances Chromatin
    Language English
    Publishing date 2022-02-08
    Publishing country United States
    Document type Introductory Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1098979-1
    ISSN 1939-4586 ; 1059-1524
    ISSN (online) 1939-4586
    ISSN 1059-1524
    DOI 10.1091/mbc.E21-08-0400
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 2853: Show issues Location:
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  2. Article: Human dynein-dynactin is a fast processive motor in living cells.

    Verma, Vikash / Wadsworth, Patricia / Maresca, Thomas J

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Minus-end directed transport along microtubules in eukaryotes is primarily mediated by cytoplasmic dynein and its cofactor dynactin. Significant advances have been made in recent years characterizing human dynein-dynactin structure and function using in ... ...

    Abstract Minus-end directed transport along microtubules in eukaryotes is primarily mediated by cytoplasmic dynein and its cofactor dynactin. Significant advances have been made in recent years characterizing human dynein-dynactin structure and function using in vitro assays, however, there is limited knowledge about the motile properties and functional organization of dynein-dynactin in living human cells. Total internal reflection fluorescence microscopy (TIRFM) of CRISPR-engineered human cells is employed here to visualize fluorescently tagged dynein heavy chain (DHC) and p50 with high spatio-temporal resolution. We find that p50 and DHC exhibit indistinguishable motility properties in their velocities, run lengths, and run times. The dynein-dynactin complexes are fast (∼1.2 μm/s) and typically run for several microns (∼2.7 μm). Quantification of the fluorescence intensities of motile puncta reveals that dynein-dynactin runs are mediated by at least one DHC dimer while the velocity is consistent with that measured for double dynein (two DHC dimers) complexes in vitro.
    Language English
    Publishing date 2023-11-29
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.11.28.569102
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  3. Article ; Online: Multimerization of a disordered kinetochore protein promotes accurate chromosome segregation by localizing a core dynein module.

    McGory, Jessica M / Verma, Vikash / Barcelos, Dylan M / Maresca, Thomas J

    The Journal of cell biology

    2024  Volume 223, Issue 3

    Abstract: Kinetochores connect chromosomes and spindle microtubules to maintain genomic integrity through cell division. Crosstalk between the minus-end directed motor dynein and kinetochore-microtubule attachment factors promotes accurate chromosome segregation ... ...

    Abstract Kinetochores connect chromosomes and spindle microtubules to maintain genomic integrity through cell division. Crosstalk between the minus-end directed motor dynein and kinetochore-microtubule attachment factors promotes accurate chromosome segregation by a poorly understood pathway. Here, we identify a linkage between the intrinsically disordered protein Spc105 (KNL1 orthologue) and dynein using an optogenetic oligomerization assay. Core pools of the checkpoint protein BubR1 and the adaptor complex RZZ contribute to the linkage. Furthermore, a minimal segment of Spc105 with a propensity to multimerize and which contains protein binding motifs is sufficient to link Spc105 to RZZ/dynein. Deletion of the minimal region from Spc105 compromises the recruitment of its binding partners to kinetochores and elevates chromosome missegregation due to merotelic attachments. Restoration of normal chromosome segregation and localization of BubR1 and RZZ requires both protein binding motifs and oligomerization of Spc105. Together, our results reveal that higher-order multimerization of Spc105 contributes to localizing a core pool of RZZ that promotes accurate chromosome segregation.
    MeSH term(s) Cell Division ; Chromosome Segregation ; Dyneins/genetics ; Intrinsically Disordered Proteins ; Kinetochores ; Drosophila/genetics ; Animals
    Chemical Substances Dyneins (EC 3.6.4.2) ; Intrinsically Disordered Proteins ; Spc105R protein, Drosophila ; BubR1 protein, Drosophila
    Language English
    Publishing date 2024-01-05
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.202211122
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  4. Article ; Online: The whole is greater than the sum of its parts: at the intersection of order, disorder, and kinetochore function.

    Audett, Margaux R / Maresca, Thomas J

    Essays in biochemistry

    2020  Volume 64, Issue 2, Page(s) 349–358

    Abstract: The kinetochore (KT) field has matured tremendously since Earnshaw first identified CENP-A, CENP-B, and CENP-C [1,2]. In the past 35 years, the accumulation of knowledge has included: defining the parts list, identifying epistatic networks of ... ...

    Abstract The kinetochore (KT) field has matured tremendously since Earnshaw first identified CENP-A, CENP-B, and CENP-C [1,2]. In the past 35 years, the accumulation of knowledge has included: defining the parts list, identifying epistatic networks of interdependence within the parts list, understanding the spatial organization of subcomplexes into a massive structure - hundreds of megadaltons in size, and dissecting the functions of the KT in its entirety as well as of its individual parts. Like nearly all cell and molecular biology fields, the structure-function paradigm has been foundational to advances in the KT field. A point nicely highlighted by the fact that we are at the precipice of the in vitro reconstitution of a functional KT holo complex. Yet conventional notions of structure cannot provide a complete picture of the KT especially since it contains an abundance of unstructured or intrinsically disordered constituents. The combination of structured and disordered proteins within the KT results in an assembled system that is functionally greater than the sum of its parts.
    MeSH term(s) Animals ; Humans ; Kinetochores/chemistry ; Kinetochores/metabolism ; Mitosis ; Spindle Apparatus
    Language English
    Publishing date 2020-10-05
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Review
    ISSN 1744-1358 ; 0071-1365
    ISSN (online) 1744-1358
    ISSN 0071-1365
    DOI 10.1042/EBC20190069
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  5. Article: An intrinsically disordered kinetochore protein coordinates mechanical regulation of chromosome segregation by dynein.

    McGory, Jessica M / Barcelos, Dylan M / Verma, Vikash / Maresca, Thomas J

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Kinetochores connect chromosomes and spindle microtubules to maintain genomic integrity through cell division. Crosstalk between the minus-end directed motor dynein and kinetochore-microtubule attachment factors promotes accurate chromosome segregation ... ...

    Abstract Kinetochores connect chromosomes and spindle microtubules to maintain genomic integrity through cell division. Crosstalk between the minus-end directed motor dynein and kinetochore-microtubule attachment factors promotes accurate chromosome segregation through a poorly understood pathway. Here we identify a physical linkage between the intrinsically disordered protein Spc105 (KNL1 orthologue) and dynein using an optogenetic oligomerization assay. Core pools of the checkpoint protein BubR1 and the adaptor complex RZZ mediate the connection of Spc105 to dynein. Furthermore, a minimal segment of Spc105 that contains regions with a propensity to multimerize and binding motifs for Bub1 and BubR1 is sufficient to functionally link Spc105 to RZZ and dynein. Deletion of the minimal region from Spc105 reduces recruitment of its binding partners to bioriented kinetochores and causes chromosome mis-segregation. Restoration of normal chromosome segregation and localization of BubR1 and RZZ requires both protein binding motifs and higher-order oligomerization of Spc105. Together, our results reveal that higher-order multimerization of Spc105 is required to recruit a core pool of RZZ that modulates microtubule attachment stability to promote accurate chromosome segregation.
    Language English
    Publishing date 2023-05-09
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.05.07.539709
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Microtubule plus-ends act as physical signaling hubs to activate RhoA during cytokinesis.

    Verma, Vikash / Maresca, Thomas J

    eLife

    2019  Volume 8

    Abstract: Microtubules (MTs) are essential for cleavage furrow positioning during cytokinesis, but the mechanisms by which MT-derived signals spatially define regions of cortical contractility are unresolved. In this study cytokinesis regulators visualized ... ...

    Abstract Microtubules (MTs) are essential for cleavage furrow positioning during cytokinesis, but the mechanisms by which MT-derived signals spatially define regions of cortical contractility are unresolved. In this study cytokinesis regulators visualized in
    MeSH term(s) Amino Acid Motifs ; Anaphase/drug effects ; Animals ; Concanavalin A/pharmacology ; Cytokinesis/drug effects ; Drosophila Proteins/metabolism ; Drosophila melanogaster/cytology ; Drosophila melanogaster/metabolism ; Green Fluorescent Proteins/metabolism ; Microtubules/drug effects ; Microtubules/metabolism ; Myosins/metabolism ; Protein Binding/drug effects ; Signal Transduction/drug effects ; rho GTP-Binding Proteins/metabolism
    Chemical Substances Drosophila Proteins ; enhanced green fluorescent protein ; Concanavalin A (11028-71-0) ; Green Fluorescent Proteins (147336-22-9) ; Myosins (EC 3.6.4.1) ; Rho1 protein, Drosophila (EC 3.6.5.2) ; rho GTP-Binding Proteins (EC 3.6.5.2)
    Language English
    Publishing date 2019-02-13
    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.38968
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  7. Article ; Online: Direct observation of branching MT nucleation in living animal cells.

    Verma, Vikash / Maresca, Thomas J

    The Journal of cell biology

    2019  Volume 218, Issue 9, Page(s) 2829–2840

    Abstract: Centrosome-mediated microtubule (MT) nucleation has been well characterized; however, numerous noncentrosomal MT nucleation mechanisms exist. The branching MT nucleation pathway envisages that the γ-tubulin ring complex (γ-TuRC) is recruited to MTs by ... ...

    Abstract Centrosome-mediated microtubule (MT) nucleation has been well characterized; however, numerous noncentrosomal MT nucleation mechanisms exist. The branching MT nucleation pathway envisages that the γ-tubulin ring complex (γ-TuRC) is recruited to MTs by the augmin complex to initiate nucleation of new MTs. While the pathway is well conserved at a molecular and functional level, branching MT nucleation by core constituents has never been directly observed in animal cells. Here, multicolor TIRF microscopy was applied to visualize and quantitatively define the entire process of branching MT nucleation in dividing
    MeSH term(s) Anaphase/physiology ; Animals ; Cytokinesis/physiology ; Drosophila Proteins/metabolism ; Drosophila melanogaster ; Microtubule-Organizing Center/metabolism ; Microtubules/metabolism ; rho GTP-Binding Proteins/metabolism
    Chemical Substances Drosophila Proteins ; Rho1 protein, Drosophila (EC 3.6.5.2) ; rho GTP-Binding Proteins (EC 3.6.5.2)
    Language English
    Publishing date 2019-07-24
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Video-Audio Media
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.201904114
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    Ub I Zs.190: Show issues Location:
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  8. Article ; Online: Cell Division: Here Comes the Kinesin Cavalry.

    Audett, Margaux R / Maresca, Thomas J

    Current biology : CB

    2018  Volume 28, Issue 17, Page(s) R943–R946

    Abstract: A new study finds that a spindle motor makes an unexpected contribution to kinetochore-microtubule attachments and chromosome segregation. ...

    Abstract A new study finds that a spindle motor makes an unexpected contribution to kinetochore-microtubule attachments and chromosome segregation.
    MeSH term(s) Cell Division ; Chromosome Segregation ; Kinesin/genetics ; Kinetochores ; Microtubules ; Protein Phosphatase 1 ; Spindle Apparatus
    Chemical Substances Protein Phosphatase 1 (EC 3.1.3.16) ; Kinesin (EC 3.6.4.4)
    Language English
    Publishing date 2018-09-11
    Publishing country England
    Document type Journal Article ; Comment
    ZDB-ID 1071731-6
    ISSN 1879-0445 ; 0960-9822
    ISSN (online) 1879-0445
    ISSN 0960-9822
    DOI 10.1016/j.cub.2018.07.054
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    Zs.A 3208: Show issues Location:
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  9. Article: Measuring mitotic forces.

    Ye, Anna A / Maresca, Thomas J

    Methods in cell biology

    2018  Volume 144, Page(s) 165–184

    Abstract: Productive chromosome movements require that a large multiprotein complex called the kinetochore assemble on sister centromeres. The kinetochore fulfills two critical functions as (1) the physical linkage between chromosomes and spindle microtubules and ( ...

    Abstract Productive chromosome movements require that a large multiprotein complex called the kinetochore assemble on sister centromeres. The kinetochore fulfills two critical functions as (1) the physical linkage between chromosomes and spindle microtubules and (2) a mechanomolecular sensor that relays a spindle assembly checkpoint signal delaying anaphase onset until chromosomes are attached to spindle microtubules and bioriented. Given its central roles in such a vital process, the kinetochore is one of the most important force-transducing structures in cells; yet it has been technically challenging to measure kinetochore forces. Barriers to measuring cellular forces have begun to be broken by the development of fluorescence-based tension sensors. In this chapter, two methods will be described for measuring kinetochore forces in living cells and strategies for applying these sensors to other force-transducing processes and molecules will be discussed.
    MeSH term(s) Animals ; Biomechanical Phenomena ; Biosensing Techniques ; Cytological Techniques/methods ; Drosophila/cytology ; Fluorescence Resonance Energy Transfer ; Mitosis ; Photobleaching ; Talin/metabolism ; Vinculin/metabolism
    Chemical Substances Talin ; Vinculin (125361-02-6)
    Language English
    Publishing date 2018-04-10
    Publishing country United States
    Document type Journal Article
    ISSN 0091-679X
    ISSN 0091-679X
    DOI 10.1016/bs.mcb.2018.03.007
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  10. Article ; Online: Microtubule plus-ends act as physical signaling hubs to activate RhoA during cytokinesis

    Vikash Verma / Thomas J Maresca

    eLife, Vol

    2019  Volume 8

    Abstract: Microtubules (MTs) are essential for cleavage furrow positioning during cytokinesis, but the mechanisms by which MT-derived signals spatially define regions of cortical contractility are unresolved. In this study cytokinesis regulators visualized in ... ...

    Abstract Microtubules (MTs) are essential for cleavage furrow positioning during cytokinesis, but the mechanisms by which MT-derived signals spatially define regions of cortical contractility are unresolved. In this study cytokinesis regulators visualized in Drosophila melanogaster (Dm) cells were found to localize to and track MT plus-ends during cytokinesis. The RhoA GEF Pebble (Dm ECT2) did not evidently tip-track, but rather localized rapidly to cortical sites contacted by MT plus-tips, resulting in RhoA activation and enrichment of myosin-regulatory light chain. The MT plus-end localization of centralspindlin was compromised following EB1 depletion, which resulted in a higher incidence of cytokinesis failure. Centralspindlin plus-tip localization depended on the C-terminus and a putative EB1-interaction motif (hxxPTxh) in RacGAP50C. We propose that MT plus-end-associated centralspindlin recruits a cortical pool of Dm ECT2 upon physical contact to activate RhoA and to trigger localized contractility.
    Keywords microtubules ; RacGAP50C ; MKLP1 ; Centralspindlin ; Polo kinase ; Cytokinesis ; Medicine ; R ; Science ; Q ; Biology (General) ; QH301-705.5
    Subject code 571
    Language English
    Publishing date 2019-02-01T00:00:00Z
    Publisher eLife Sciences Publications Ltd
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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