Mitotic Chromosome Mechanics: How Cells Segregate Their Genome

Batty, Paul; Gerlich, Daniel W.

doi:10.1016/j.tcb.2019.05.007

Cited by 98 publications

(82 citation statements)

References 98 publications

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“…Within interphase nuclei, Ki67 is also important in establishing the boundary between nucleoli and surrounding heterochromatin; for recent discussions of the multiple roles of Ki67, see [33,34]. Disruption of the interaction between Ki67 and mitotic chromosome arms, employing siRNA [32,35] or auxin-induced 'degron' degradation [33] of Ki67, results in a collapse of mitotic chromosomes. We wanted to explore whether the hyperosmotic sucrose stress-induced congelation of mitotic chromosomes influences the distribution of Ki67 in separating the mitotic chromosome arms.…”

Section: Ki67 Segregates From Hyperosmotic Sucrose Congealed Mitotic mentioning

confidence: 99%

Hyperosmotic stress: in situ chromatin phase separation

Olins

Gould

Boyd

et al. 2020

Nucleus

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Dehydration of cells by acute hyperosmotic stress has profound effects upon cell structure and function. Interphase chromatin and mitotic chromosomes collapse ("congelation"). HL-60/S4 cells remain~100% viable for, at least, 1 hour, exhibiting shrinkage to~2/3 their original volume, when placed in 300mM sucrose in tissue culture medium. Fixed cells were imaged by immunostaining confocal and STED microscopy. At a "global" structural level (μm), mitotic chromosomes congeal into a residual gel with apparent (phase) separations of Ki67, CTCF, SMC2, RAD21, H1 histones and HMG proteins. At an "intermediate" level (sub-μm), radial distribution analysis of STED images revealed a most probable peak DNA density separation of~0.16 μm, essentially unchanged by hyperosmotic stress. At a "local" structural level (~1-2 nm), in vivo crosslinking revealed essentially unchanged crosslinked products between H1, HMG and inner histones. Hyperosmotic cellular stress is discussed in terms of concepts of mitotic chromosome structure and liquid-liquid phase separation.

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Section: Ki67 Segregates From Hyperosmotic Sucrose Congealed Mitotic mentioning

confidence: 99%

Hyperosmotic stress: in situ chromatin phase separation

Olins

Gould

Boyd

et al. 2020

Nucleus

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“…C21orf45, also known as MIS18A, is a kinetocore protein belonging to the Mis18 complex assembly that is crucial for CENPA deposition at the centromere [26,27]. DEPDC1 is highly expressed in M phase and its silencing caused mitotic defects [28][29][30][31] and NCAPH is a regulatory subunit of the condensin complex required for the conversion of interphase chromatin into condensed chromosomes [32][33][34]. Overall, their altered expression could participate in the definition of the aberrant mitotic regulation.…”

Section: P53 Mut Downstream Targets Are Differently Modulated In Pt-rmentioning

confidence: 99%

Identification and Characterization of a New Platinum-Induced TP53 Mutation in MDAH Ovarian Cancer Cells

Lorenzon

Pellarin

Pellizzari

et al. 2019

Cells

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Platinum-based chemotherapy is the therapy of choice for epithelial ovarian cancer (EOC). Acquired resistance to platinum (PT) is a frequent event that leads to disease progression and predicts poor prognosis. To understand possible mechanisms underlying acquired PT-resistance, we have recently generated and characterized three PT-resistant isogenic EOC cell lines. Here, we more deeply characterize several PT-resistant clones derived from MDAH-2774 cells. We show that, in these cells, the increased PT resistance was accompanied by the presence of a subpopulation of multinucleated giant cells. This phenotype was likely due to an altered progression through the M phase of the cell cycle and accompanied by the deregulated expression of genes involved in M phase progression known to be target of mutant TP53. Interestingly, we found that PT-resistant MDAH cells acquired in the TP53 gene a novel secondary mutation (i.e., S185G) that accompanied the R273H typical of MDAH cells. The double p53 S185G/R273H mutant increases the resistance to PT in a TP53 null EOC cellular model. Overall, we show how the selective pressure of PT is able to induce additional mutation in an already mutant TP53 gene in EOC and how this event could contribute to the acquisition of novel cellular phenotypes.Cells 2020, 9, 36 2 of 18 PT resistance has been linked to alterations in several processes such as drug transport, drug inactivation, DNA damage response, DNA repair, apoptosis, and autophagy [8][9][10][11]. Many studies have been done to identify genes and mechanisms directly associated to resistance to PT therapy. We have recently contributed to this issue reporting the selection and preliminary characterization of three new isogenic models of PT-resistant EOC cell lines-MDAH-2774, TOV-112D and OVSAHO. This work has pointed out a common ability of the three isogenic PT-resistant cells to resolve the PT-induced DNA damage compared to parental cells. Moreover, all PT-resistant cells displayed a change in their morphology and a higher ability to grow on mesothelium [12].In the present work, we better characterized MDAH-2774 PT-resistant (PT-res) cells reporting the appearance of a novel mutation in TP53 induced by platinum pressure that is linked to the increased resistance to PT and an altered mitotic division observed in PT-res cells. Materials and Methods Cell CultureMDAH-2774 (CRL-10303) parental and SKOV-3 (HTB-77) cells are from ATCC. Cisplatin-resistant (PT-res) isogenic cells were generated as described [12]. All cell lines were maintained in RPMI-1640 medium (Sigma-Aldrich Co., St. Louis, MO, USA) containing 10% heat-inactivated FBS, 100 U/mL penicillin and streptomycin (Sigma-Aldrich Co., St. Louis, MO, USA) at 37 • C in a 5% CO 2 atmosphere. After the selection process, PT-res cells were kept in PT-free medium. MDAH-2774 PT-res clones were obtained from PT-res pools by plating them at 0.7 cell/well dilution in a 96-multiwell plate. All cell lines were authenticated in our lab using the Cell ID TM System (Promega, Madison, WI, USA) ...

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“…Thus, we wondered if we can visualize this interaction in a living cell. Chromosomes are present as a contiguous mass in the interphase nuclei and appear as distinct bodies during late prophase (reviewed in Batty and Gerlich, 2019). Therefore, we sought to examine the localization of NuMA in synchronized non-transformed hTERT-RPE1 cells during late prophase before NEBD ( Fig.…”

Section: Cdk1-mediated Phosphorylation Release Numa From Chromatin Atmentioning

confidence: 99%

NuMA interaction with chromatin is vital for proper nuclear architecture in human cells

Rajeevan¹,

Keshri²,

Kotak³

2020

Preprint

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INDIAPh: +91 80 2293 2292 sachinkotak@iisc.ac.in NuMA regulates nuclear shape independent of its mitotic function Abstract NuMA is an abundant long-coiled-coil protein that plays a prominent role in spindle organization during mitosis. In interphase, NuMA is localized to the nucleus and hypothesized to control gene expression and chromatin organization. However, because of the prominent mitotic phenotype upon NuMA loss, its precise function in the interphase nucleus remains elusive. Here, we report that NuMA is associated with chromatin in interphase and prophase but released upon nuclear envelope breakdown (NEBD) by the action of Cdk1. We uncovered that NuMA directly interacts with DNA via evolutionarily conserved sequences in its C-terminus. Notably, the expression of the DNA-binding mutant of NuMA affects chromatin decondensation at the mitotic exit, and nuclear shape in interphase. The impact on nuclear shape by mutant NuMA expression is due to its potential to polymerize into high-order fibrillar structures. This study links the chromatin binding ability of NuMA with the maintenance of nuclear shape and architecture, which has a well-studied role in regulating gene expression during development and diseases. localizes at the spindle poles and cell cortex, where it is required for the proper assembly and maintenance of the mitotic spindle as well as spindle orientation and elongation (Lydersen and Pettijohn

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Mitotic Chromosome Mechanics: How Cells Segregate Their Genome

Cited by 98 publications

References 98 publications

Hyperosmotic stress: in situ chromatin phase separation

Hyperosmotic stress: in situ chromatin phase separation

Identification and Characterization of a New Platinum-Induced TP53 Mutation in MDAH Ovarian Cancer Cells

NuMA interaction with chromatin is vital for proper nuclear architecture in human cells

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