scholarly journals Kinesin-binding protein ensures accurate chromosome segregation by buffering KIF18A and KIF15

2019 ◽  
Vol 218 (4) ◽  
pp. 1218-1234 ◽  
Author(s):  
Heidi L.H. Malaby ◽  
Megan E. Dumas ◽  
Ryoma Ohi ◽  
Jason Stumpff
Keyword(s):  
Motor Activity ◽  
Chromosome Segregation ◽  
Binding Protein ◽  
Mitotic Chromosomes ◽  
Microtubule Binding ◽  
Daughter Cells ◽  
Segregation Of Chromosomes ◽  
In Cells

Mitotic kinesins must be regulated to ensure a precise balance of spindle forces and accurate segregation of chromosomes into daughter cells. Here, we demonstrate that kinesin-binding protein (KBP) reduces the activity of KIF18A and KIF15 during metaphase. Overexpression of KBP disrupts the movement and alignment of mitotic chromosomes and decreases spindle length, a combination of phenotypes observed in cells deficient for KIF18A and KIF15, respectively. We show through gliding filament and microtubule co-pelleting assays that KBP directly inhibits KIF18A and KIF15 motor activity by preventing microtubule binding. Consistent with these effects, the mitotic localizations of KIF18A and KIF15 are altered by overexpression of KBP. Cells depleted of KBP exhibit lagging chromosomes in anaphase, an effect that is recapitulated by KIF15 and KIF18A overexpression. Based on these data, we propose a model in which KBP acts as a protein buffer in mitosis, protecting cells from excessive KIF18A and KIF15 activity to promote accurate chromosome segregation.

scholarly journals Kinesin-Binding Protein (KBP) buffers the activity of KIF18A and KIF15 in mitosis to ensure accurate chromosome segregation

2018 ◽  
Author(s):  
Heidi L. H. Malaby ◽  
Megan E. Dumas ◽  
Ryoma Ohi ◽  
Jason Stumpff
Keyword(s):  
Motor Activity ◽  
Chromosome Segregation ◽  
Binding Protein ◽  
Mitotic Chromosomes ◽  
Microtubule Binding ◽  
Daughter Cells ◽  
Chromosome Alignment ◽  
Segregation Of Chromosomes ◽  
In Cells

ABSTRACTMitotic kinesins must be regulated to ensure a precise balance of spindle forces and accurate segregation of chromosomes into daughter cells. Here we demonstrate that Kinesin-Binding Protein (KBP) reduces the activity of KIF18A and KIF15 during metaphase. Overexpression of KBP disrupts the movement and alignment of mitotic chromosomes and decreases spindle length, a combination of phenotypes observed in cells deficient for KIF18A and KIF15, respectively. We show through gliding filament and microtubule co-pelleting assays that KBP directly inhibits KIF18A and KIF15 motor activity by preventing microtubule-binding. Consistent with these effects, the mitotic localizations of KIF18A and KIF15 are altered by overexpression of KBP. Cells depleted of KBP exhibit lagging chromosomes in anaphase, an effect that is recapitulated by KIF15 and KIF18A overexpression. Based on these data, we propose a model in which KBP acts as a protein buffer in mitosis, protecting cells from excessive KIF18A and KIF15 activity to promote accurate chromosome segregation.SUMMARYKinesin-Binding Protein (KBP) is identified as a regulator of the kinesins KIF18A and KIF15 during mitosis. KBP buffers the activity of these motors to control chromosome alignment and spindle integrity in metaphase and prevent lagging chromosomes in anaphase.

scholarly journals Whole-proteome genetic analysis of dependencies in assembly of a vertebrate kinetochore

2015 ◽  
Vol 211 (6) ◽  
pp. 1141-1156 ◽  
Author(s):  
Itaru Samejima ◽  
Christos Spanos ◽  
Flavia de Lima Alves ◽  
Tetsuya Hori ◽  
Marinela Perpelescu ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Mitotic Chromosome ◽  
Protein Complexes ◽  
Interacting Proteins ◽  
Quantitative Mass Spectrometry ◽  
Mitotic Chromosomes ◽  
Microtubule Binding ◽  
Checkpoint Signaling ◽  
Ndc80 Complex ◽  
Stable Association

Kinetochores orchestrate mitotic chromosome segregation. Here, we use quantitative mass spectrometry of mitotic chromosomes isolated from a comprehensive set of chicken DT40 mutants to examine the dependencies of 93 confirmed and putative kinetochore proteins for stable association with chromosomes. Clustering and network analysis reveal both known and unexpected aspects of coordinated behavior for members of kinetochore protein complexes. Surprisingly, CENP-T depends on CENP-N for chromosome localization. The Ndc80 complex exhibits robust correlations with all other complexes in a “core” kinetochore network. Ndc80 associated with CENP-T interacts with a cohort of Rod, zw10, and zwilch (RZZ)–interacting proteins that includes Spindly, Mad1, and CENP-E. This complex may coordinate microtubule binding with checkpoint signaling. Ndc80 associated with CENP-C forms the KMN (Knl1, Mis12, Ndc80) network and may be the microtubule-binding “workhorse” of the kinetochore. Our data also suggest that CENP-O and CENP-R may regulate the size of the inner kinetochore without influencing the assembly of the outer kinetochore.

scholarly journals PinX1 Is a Novel Microtubule-binding Protein Essential for Accurate Chromosome Segregation

2009 ◽  
Vol 284 (34) ◽  
pp. 23072-23082 ◽  
Author(s):  
Kai Yuan ◽  
Na Li ◽  
Kai Jiang ◽  
Tongge Zhu ◽  
Yuda Huo ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Binding Protein ◽  
Microtubule Binding

scholarly journals PICH promotes SUMOylated TopoisomeraseIIα dissociation from mitotic centromeres for proper chromosome segregation

2019 ◽  
Author(s):  
Victoria Hassebroek ◽  
Hyewon Park ◽  
Nootan Pandey ◽  
Brooklyn T. Lerbakken ◽  
Vasilisa Aksenova ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Binding Protein ◽  
Binding Activity ◽  
Mitotic Chromosomes ◽  
Summary Statement ◽  
Dna Translocase

AbstractPolo-like kinase interacting checkpoint helicase (PICH) is a SNF2 family DNA translocase and is a Small Ubiquitin-like modifier (SUMO) binding protein. Despite that both translocase activity and SUMO-binding activity are required for proper chromosome segregation, how these two activities function to mediate chromosome segregation remains unknown. Here, we show that PICH specifically promotes dissociation of SUMOylated TopoisomeraseIIα (TopoIIα) from mitotic chromosomes. When TopoIIα is stalled by treatment of cells with a potent TopoII inhibitor, ICRF-193, TopoIIα becomes SUMOylated, and this promotes its interaction with PICH. Conditional depletion of PICH using the Auxin Inducible Degron (AID) system resulted in retention of SUMOylated TopoIIα on chromosomes, indicating that PICH removes stalled SUMOylated TopoIIα from chromosomes. In vitro assays showed that PICH specifically regulates SUMOylated TopoIIα activity using its SUMO-binding and translocase activities. Taken together, we propose a novel mechanism for how PICH acts on stalled SUMOylated TopoIIα for proper chromosome segregation.Summary StatementPolo-like kinase interacting checkpoint helicase (PICH) interacts with SUMOylated proteins to mediate proper chromosome segregation during mitosis. The results demonstrate that PICH promotes dissociation of SUMOylated TopoisomeraseIIα from chromosomes and that function leads to proper chromosome segregation.

scholarly journals Centromeric Non-Coding RNAs: Conservation and Diversity in Function

2020 ◽  
Vol 6 (1) ◽  
pp. 4
Author(s):  
Takashi Ideue ◽  
Tokio Tani
Keyword(s):  
Chromosome Segregation ◽  
Genetic Material ◽  
Experimental Studies ◽  
Mitotic Chromosomes ◽  
Daughter Cells ◽  
Non Coding Rnas ◽  
Available Information

Chromosome segregation is strictly regulated for the proper distribution of genetic material to daughter cells. During this process, mitotic chromosomes are pulled to both poles by bundles of microtubules attached to kinetochores that are assembled on the chromosomes. Centromeres are specific regions where kinetochores assemble. Although these regions were previously considered to be silent, some experimental studies have demonstrated that transcription occurs in these regions to generate non-coding RNAs (ncRNAs). These centromeric ncRNAs (cenRNAs) are involved in centromere functions. Here, we describe the currently available information on the functions of cenRNAs in several species.

scholarly journals A Stu2-mediated intrinsic tension-sensing pathway promotes chromosome biorientation in vivo

2018 ◽  
Author(s):  
Matthew P. Miller ◽  
Rena K. Evans ◽  
Alex Zelter ◽  
Elisabeth A. Geyer ◽  
Michael J. MacCoss ◽  
...  
Keyword(s):  
Error Correction ◽  
Chromosome Segregation ◽  
Aurora B ◽  
Critical Aspect ◽  
Daughter Cells ◽  
Low Tension ◽  
Kinetochore Function ◽  
Segregation Of Chromosomes

ABSTRACTAccurate segregation of chromosomes to daughter cells is a critical aspect of cell division. It requires the kinetochores on duplicated chromosomes to biorient, attaching to microtubules from opposite poles of the cell. Bioriented attachments come under tension, while incorrect attachments lack tension and must be destabilized. A well-studied error correction pathway is mediated by the Aurora B kinase, which destabilizes low tension-bearing attachments. We recently discovered that in vitro, kinetochores display an additional intrinsic tension-sensing pathway that utilizes Stu2. This pathway’s contribution to error correction in cells, however, was unknown. Here, we identify a Stu2 mutant that abolishes its kinetochore function and show that it causes error correction defects in vivo. We also show that this intrinsic tension-sensing pathway functions in concert with the Aurora B-mediated pathway. Together, our work indicates that cells employ at least two pathways to ensure biorientation and the accuracy of chromosome segregation.

scholarly journals ALIX and ESCRT-I/II function as parallel ESCRT-III recruiters in cytokinetic abscission

2016 ◽  
Vol 212 (5) ◽  
pp. 499-513 ◽  
Author(s):  
Liliane Christ ◽  
Eva M. Wenzel ◽  
Knut Liestøl ◽  
Camilla Raiborg ◽  
Coen Campsteijn ◽  
...  
Keyword(s):  
Cell Division ◽  
Final Stage ◽  
Binding Protein ◽  
Intercellular Bridge ◽  
Checkpoint Signaling ◽  
Endosomal Sorting ◽  
Daughter Cells ◽  
Escrt Machinery ◽  
Chromosome Bridges ◽  
In Cells

Cytokinetic abscission, the final stage of cell division where the two daughter cells are separated, is mediated by the endosomal sorting complex required for transport (ESCRT) machinery. The ESCRT-III subunit CHMP4B is a key effector in abscission, whereas its paralogue, CHMP4C, is a component in the abscission checkpoint that delays abscission until chromatin is cleared from the intercellular bridge. How recruitment of these components is mediated during cytokinesis remains poorly understood, although the ESCRT-binding protein ALIX has been implicated. Here, we show that ESCRT-II and the ESCRT-II–binding ESCRT-III subunit CHMP6 cooperate with ESCRT-I to recruit CHMP4B, with ALIX providing a parallel recruitment arm. In contrast to CHMP4B, we find that recruitment of CHMP4C relies predominantly on ALIX. Accordingly, ALIX depletion leads to furrow regression in cells with chromosome bridges, a phenotype associated with abscission checkpoint signaling failure. Collectively, our work reveals a two-pronged recruitment of ESCRT-III to the cytokinetic bridge and implicates ALIX in abscission checkpoint signaling.

scholarly journals A quantitative and semi-automated method for determining misaligned and lagging chromosomes during mitosis

2020 ◽  
pp. mbc.E20-09-0585
Author(s):  
Luciano Gama Braga ◽  
Diogjena Katerina Prifti ◽  
Chantal Garand ◽  
Pawan Kumar Saini ◽  
Sabine Elowe
Keyword(s):  
Chromosome Segregation ◽  
Hallmarks Of Cancer ◽  
Analytical Geometry ◽  
Daughter Cells ◽  
Chromosome Missegregation ◽  
Automated Method ◽  
Sister Chromatids ◽  
Chromosome Congression ◽  
Chromosome Alignment ◽  
In Cells

Accurate chromosome alignment at metaphase facilitates the equal segregation of sister chromatids to each of the nascent daughter cells. Lack of proper metaphase alignment is an indicator of defective chromosome congression and aberrant kinetochore-microtubule attachments which in turn promotes chromosome missegregation and aneuploidy, hallmarks of cancer. Tools to sensitively, accurately and quantitatively measure chromosome alignment at metaphase will facilitate understanding of the contribution of chromosome segregation errors to the development of aneuploidy. In this work, we have developed and validated a method based on analytical geometry to measure several indicators of chromosome misalignment. We generated semi-automated and flexible ImageJ2/Fiji pipelines to quantify kinetochore misalignment at metaphase plates as well as lagging chromosomes at anaphase. These tools will ultimately allow sensitive and systematic quantitation of these chromosome segregation defects in cells undergoing mitosis.

scholarly journals The kinetic landscape of an RNA-binding protein in cells

Nature ◽  
2021 ◽  
Vol 591 (7848) ◽  
pp. 152-156 ◽  
Author(s):  
Deepak Sharma ◽  
Leah L. Zagore ◽  
Matthew M. Brister ◽  
Xuan Ye ◽  
Carlos E. Crespo-Hernández ◽  
...  
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
In Cells

scholarly journals Identification of a novel nucleolar protein complex required for mitotic chromosome segregation through centromeric accumulation of Aurora B

2020 ◽  
Vol 48 (12) ◽  
pp. 6583-6596
Author(s):  
Akiko Fujimura ◽  
Yuki Hayashi ◽  
Kazashi Kato ◽  
Yuichiro Kogure ◽  
Mutsuro Kameyama ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Protein Complex ◽  
Metaphase Plate ◽  
Nucleolar Protein ◽  
Aurora B ◽  
Mitotic Progression ◽  
Mitotic Chromosomes ◽  
Nucleolar Proteins ◽  
Chromatid Cohesion ◽  
Wd Repeat

Abstract The nucleolus is a membrane-less nuclear structure that disassembles when cells undergo mitosis. During mitosis, nucleolar factors are thus released from the nucleolus and dynamically change their subcellular localization; however, their functions remain largely uncharacterised. Here, we found that a nucleolar factor called nucleolar protein 11 (NOL11) forms a protein complex with two tryptophan-aspartic acid (WD) repeat proteins named WD-repeat protein 43 (WDR43) and Cirhin in mitotic cells. This complex, referred to here as the NWC (NOL11-WDR43-Cirhin) complex, exists in nucleoli during interphase and translocates to the periphery of mitotic chromosomes, i.e., perichromosomal regions. During mitotic progression, both the congression of chromosomes to the metaphase plate and sister chromatid cohesion are impaired in the absence of the NWC complex, as it is required for the centromeric enrichment of Aurora B and the associating phosphorylation of histone H3 at threonine 3. These results reveal the characteristics of a novel protein complex consisting of nucleolar proteins, which is required for regulating kinetochores and centromeres to ensure faithful chromosome segregation.

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