bims-micesi 2022-07-03 papers

bims-micesi

Biomed News

on Mitotic cell signalling

Issue of 2022‒07‒03
eleven papers selected by
Valentina Piano
Max Planck Institute of Molecular Physiology

BubR1 recruitment to the kinetochore via Bub1 enhances Spindle Assembly Checkpoint signaling.
[Getting in shape to divide: When oncogenes contribute to mitotic cell rounding].
Maintaining soluble protein homeostasis between nuclear and cytoplasmic compartments across mitosis.
Absence of a robust mitotic timer mechanism in early preimplantation mouse embryos leads to chromosome instability.
Fine-tuning cell organelle dynamics during mitosis by small GTPases.
Cell cycle-dependent binding between Cyclin B1 and Cdk1 revealed by time-resolved fluorescence correlation spectroscopy.
Piperine Increases Pentagamavunon-1 Anti-cancer Activity on 4T1 Breast Cancer Through Mitotic Catastrophe Mechanism and Senescence with Sharing Targeting on Mitotic Regulatory Proteins.
Dissecting the roles of Haspin and VRK1 in histone H3 phosphorylation during mitosis.
Mechanistic basis for Sgo1-mediated centromere localization and function of the CPC.
How Microtubules Build the Spindle Branch by Branch.
The impact of monosomies, trisomies and segmental aneuploidies on chromosomal stability.

Mol Biol Cell. 2022 Jun 29. mbcE22030085

BubR1 recruitment to the kinetochore via Bub1 enhances Spindle Assembly Checkpoint signaling.

Anand Banerjee, Chu Chen, Lauren Humphrey, John J Tyson, Ajit P Joglekar.

During mitosis, unattached kinetochores in a dividing cell activate the Spindle Assembly Checkpoint (SAC) and delay anaphase onset by generating the anaphase-inhibitory Mitotic Checkpoint Complex (MCC). These kinetochores generate the MCC by recruiting its constituent proteins, including BubR1. In principle, BubR1 recruitment to signaling kinetochores should increase its local concentration and promote MCC formation. However, in human cells BubR1 is mainly thought to sensitize the SAC to silencing. Whether BubR1 localization to signaling kinetochores by itself enhances SAC signaling remains unknown. Therefore, we used ectopic SAC activation systems (eSACs) to isolate two molecules that recruit BubR1 to the kinetochore: the checkpoint protein Bub1 and the KI and MELT motifs in the kinetochore protein KNL1 and observed their contribution to eSAC signaling. Our quantitative analyses and mathematical modeling show that Bub1-mediated BubR1 recruitment to the human kinetochore promotes SAC signaling and highlight BubR1's dual role of strengthening the SAC directly and silencing it indirectly. [Media: see text] [Media: see text].

DOI: https://doi.org/10.1091/mbc.E22-03-0085
Med Sci (Paris). 2022 Jun-Jul;38(6-7):38(6-7): 514-516

[Getting in shape to divide: When oncogenes contribute to mitotic cell rounding].

Danièle Salaün, Pascal Verdier-Pinard, Ali Badache.

DOI: https://doi.org/10.1051/medsci/2022066
Trends Cell Biol. 2022 Jun 28. pii: S0962-8924(22)00141-6. [Epub ahead of print]

Maintaining soluble protein homeostasis between nuclear and cytoplasmic compartments across mitosis.

Sabina Y van der Zanden, Marlieke L M Jongsma, Anna C M Neefjes, Ilana Berlin, Jacques Neefjes.

  The nuclear envelope (NE) is central to the architecture of eukaryotic cells, both as a physical barrier separating the nucleus from the cytoplasm and as gatekeeper of selective transport between them. However, in open mitosis, the NE fragments to allow for spindle formation and segregation of chromosomes, resulting in intermixing of nuclear and cytoplasmic soluble fractions. Recent studies have shed new light on the mechanisms driving reinstatement of soluble proteome homeostasis following NE reformation in daughter cells. Here, we provide an overview of how mitotic cells confront this challenge to ensure continuity of basic cellular functions across generations and elaborate on the implications for the proteasome - a macromolecular machine that functions in both cytoplasmic and nuclear compartments.

Keywords:  chromatin condensation; mitosis; nuclear envelope; proteasome; protein homeostasis

DOI:  https://doi.org/10.1016/j.tcb.2022.06.002
Development. 2022 Jul 01. pii: dev200391. [Epub ahead of print]149(13):

Absence of a robust mitotic timer mechanism in early preimplantation mouse embryos leads to chromosome instability.

Adélaïde Allais, Greg FitzHarris.

  Preimplantation embryos often consist of a combination of euploid and aneuploid cells, suggesting that safeguards preventing the generation and propagation of aneuploid cells in somatic cells might be deficient in embryos. In somatic cells, a mitotic timer mechanism has been described, in which even a small increase in the duration of M phase can cause a cell cycle arrest in the subsequent interphase, preventing further propagation of cells that have undergone a potentially hazardously long M phase. Here, we report that cell divisions in the mouse embryo and embryonic development continue even after a mitotic prolongation of several hours. However, similar M-phase extensions caused cohesion fatigue, resulting in prematurely separated sister chromatids and the production of micronuclei. Only extreme prolongation of M phase caused a subsequent interphase arrest, through a mechanism involving DNA damage. Our data suggest that the simultaneous absence of a robust mitotic timer and susceptibility of the embryo to cohesion fatigue could contribute to chromosome instability in mammalian embryos. This article has an associated 'The people behind the papers' interview.

Keywords:  Aneuploidy; Cohesion fatigue; DNA damage; M phase prolongation

DOI:  https://doi.org/10.1242/dev.200391
Front Med. 2022 Jun 27.

Fine-tuning cell organelle dynamics during mitosis by small GTPases.

Zijian Zhang, Wei Zhang, Quentin Liu.

  During mitosis, the allocation of genetic material concurs with organelle transformation and distribution. The coordination of genetic material inheritance with organelle dynamics directs accurate mitotic progression, cell fate determination, and organismal homeostasis. Small GTPases belonging to the Ras superfamily regulate various cell organelles during division. Being the key regulators of membrane dynamics, the dysregulation of small GTPases is widely associated with cell organelle disruption in neoplastic and non-neoplastic diseases, such as cancer and Alzheimer's disease. Recent discoveries shed light on the molecular properties of small GTPases as sophisticated modulators of a remarkably complex and perfect adaptors for rapid structure reformation. This review collects current knowledge on small GTPases in the regulation of cell organelles during mitosis and highlights the mediator role of small GTPase in transducing cell cycle signaling to organelle dynamics during mitosis.

Keywords:  cell organelle; mitosis; small GTPase

DOI:  https://doi.org/10.1007/s11684-022-0926-1
Open Biol. 2022 Jun;12(6): 220057

Cell cycle-dependent binding between Cyclin B1 and Cdk1 revealed by time-resolved fluorescence correlation spectroscopy.

Martina Barbiero, Luca Cirillo, Sapthaswaran Veerapathiran, Catherine Coates, Camilla Ruffilli, Jonathon Pines.

  Measuring the dynamics with which the regulatory complexes assemble and disassemble is a crucial barrier to our understanding of how the cell cycle is controlled that until now has been difficult to address. This considerable gap in our understanding is due to the difficulty of reconciling biochemical assays with single cell-based techniques, but recent advances in microscopy and gene editing techniques now enable the measurement of the kinetics of protein-protein interaction in living cells. Here, we apply fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy to study the dynamics of the cell cycle machinery, beginning with Cyclin B1 and its binding to its partner kinase Cdk1 that together form the major mitotic kinase. Although Cyclin B1 and Cdk1 are known to bind with high affinity, our results reveal that in living cells there is a pool of Cyclin B1 that is not bound to Cdk1. Furthermore, we provide evidence that the affinity of Cyclin B1 for Cdk1 increases during the cell cycle, indicating that the assembly of the complex is a regulated step. Our work lays the groundwork for studying the kinetics of protein complex assembly and disassembly during the cell cycle in living cells.

Keywords:  Cdk1; FCS; cell cycle; complex assembly; cyclin

DOI:  https://doi.org/10.1098/rsob.220057
Iran J Pharm Res. 2022 Dec;21(1): e123820

Piperine Increases Pentagamavunon-1 Anti-cancer Activity on 4T1 Breast Cancer Through Mitotic Catastrophe Mechanism and Senescence with Sharing Targeting on Mitotic Regulatory Proteins.

Endah Endah, Febri Wulandari, Yurananda Putri, Riris Istighfari Jenie, Edy Meiyanto.

  Pentagamavunon-1 performs more potent anti-cancer effects than curcumin against various cancer cells, but it remains to be optimized. Piperine shows the activity as an enhancer of a therapeutic agent. This study expects to achieve higher effectiveness of PGV-1 on 4T1 breast cancer cells through co-treatment with piperine with exploring the effect of cytotoxicity, mitotic catastrophe, cellular senescence, and target proteins of PGV-1 and piperine on the regulation of mitosis in TNBC cells (4T1). The assays emphasize MTT assay, May Grünwald-Giemsa staining, SA-β-galactosidase assay, and bioinformatics analysis, respectively, to elicit the respected activities. The results revealed that PGV-1 performed a cytotoxic effect with an IC50 value of 9 µM while piperine showed a lower cytotoxic effect with an IC50 value of 800 µM on 4T1 cells 24 h treatment. However, the combination treatment of both showed a synergistic cytotoxic enhancement effect with an average CI value < 1. Furthermore, the combination of PGV-1 and piperine induced mitotic catastrophe and senescence better than the single treatment. Treatment of 1 µM of PGV-1 and 400 µM of piperine increased the percentage of senescent cells by 33%. Bioinformatics analysis revealed that PGV-1 and piperine target proteins play a role in mitotic regulation, namely CDK1, KIF11, AURKA, AURKB, and PLK1, to contribute to mitotic catastrophe. Therefore, piperine increases the effectiveness of PGV-1 to suppress 4T1 cells growth synergistically that may occur through mitotic catastrophe and senescence targeting on mitotic regulatory proteins.

Keywords:  4T1 Cells; Mitotic Catastrophe; Pentagamavunon-1 (PGV-1); Piperine; Protein Target; Senescence; TNBC

DOI:  https://doi.org/10.5812/ijpr.123820
Sci Rep. 2022 Jul 01. 12(1): 11210

Dissecting the roles of Haspin and VRK1 in histone H3 phosphorylation during mitosis.

Tyrell N Cartwright, Rebecca J Harris, Stephanie K Meyer, Aye M Mon, Nikolaus A Watson, Cheryl Tan, Agathe Marcelot, Fangwei Wang, Sophie Zinn-Justin, Paula Traktman, Jonathan M G Higgins.

Protein kinases that phosphorylate histones are ideally-placed to influence the behavior of chromosomes during cell division. Indeed, a number of conserved histone phosphorylation events occur prominently during mitosis and meiosis in most eukaryotes, including on histone H3 at threonine-3 (H3T3ph). At least two kinases, Haspin and VRK1 (NHK-1/ballchen in Drosophila), have been proposed to carry out this modification. Phosphorylation of H3 by Haspin has defined roles in mitosis, but the significance of VRK1 activity towards histones in dividing cells has been unclear. Here, using in vitro kinase assays, KiPIK screening, RNA interference, and CRISPR/Cas9 approaches, we were unable to substantiate a direct role for VRK1, or its paralogue VRK2, in the phosphorylation of threonine-3 or serine-10 of Histone H3 in mitosis, although loss of VRK1 did slow cell proliferation. We conclude that the role of VRKs, and their more recently identified association with neuromuscular disease and importance in cancers of the nervous system, are unlikely to involve mitotic histone kinase activity. In contrast, Haspin is required to generate H3T3ph during mitosis.

DOI: https://doi.org/10.1038/s41598-022-15339-8
J Cell Biol. 2022 Aug 01. pii: e202108156. [Epub ahead of print]221(8):

Mechanistic basis for Sgo1-mediated centromere localization and function of the CPC.

Maria Alba Abad, Tanmay Gupta, Michael A Hadders, Amanda Meppelink, J Pepijn Wopken, Elizabeth Blackburn, Juan Zou, Anjitha Gireesh, Lana Buzuk, David A Kelly, Toni McHugh, Juri Rappsilber, Susanne M A Lens, A Arockia Jeyaprakash.

Centromere association of the chromosomal passenger complex (CPC; Borealin-Survivin-INCENP-Aurora B) and Sgo1 is crucial for chromosome biorientation, a process essential for error-free chromosome segregation. Phosphorylated histone H3 Thr3 (H3T3ph; directly recognized by Survivin) and histone H2A Thr120 (H2AT120ph; indirectly recognized via Sgo1), together with CPC's intrinsic nucleosome-binding ability, facilitate CPC centromere recruitment. However, the molecular basis for CPC-Sgo1 binding and how their physical interaction influences CPC centromere localization are lacking. Here, using an integrative structure-function approach, we show that the "histone H3-like" Sgo1 N-terminal tail-Survivin BIR domain interaction acts as a hotspot essential for CPC-Sgo1 assembly, while downstream Sgo1 residues and Borealin contribute for high-affinity binding. Disrupting Sgo1-Survivin interaction abolished CPC-Sgo1 assembly and perturbed CPC centromere localization and function. Our findings reveal that Sgo1 and H3T3ph use the same surface on Survivin to bind CPC. Hence, it is likely that these interactions take place in a spatiotemporally restricted manner, providing a rationale for the Sgo1-mediated "kinetochore-proximal" CPC centromere pool.

DOI: https://doi.org/10.1083/jcb.202108156
Annu Rev Cell Dev Biol. 2022 Jun 27.

How Microtubules Build the Spindle Branch by Branch.

Sophie M Travis, Brian P Mahon, Sabine Petry.

The microtubule (MT) cytoskeleton provides the architecture that governs intracellular organization and the regulated motion of macromolecules through the crowded cytoplasm. The key to establishing a functioning cytoskeletal architecture is regulating when and where new MTs are nucleated. Within the spindle, the vast majority of MTs are generated through a pathway known as branching MT nucleation, which exponentially amplifies MT number in a polar manner. Whereas other MT nucleation pathways generally require a complex organelle such as the centrosome or Golgi apparatus to localize nucleation factors, the branching site is based solely on a simple, preformed MT, making it an ideal system to study MT nucleation. In this review, we address recent developments in characterizing branching factors, the branching reaction, and its regulation, as well as branching MT nucleation in systems beyond the spindle and within human disease. Expected final online publication date for the Annual Review of Cell and Developmental Biology Volume 38 is October 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

DOI: https://doi.org/10.1146/annurev-cellbio-120420-114559
PLoS One. 2022 ;17(7): e0268579

The impact of monosomies, trisomies and segmental aneuploidies on chromosomal stability.

Dorine C Hintzen, Mar Soto, Michael Schubert, Bjorn Bakker, Diana C J Spierings, Karoly Szuhai, Peter M Lansdorp, Roel J C Kluin, Floris Foijer, René H Medema, Jonne A Raaijmakers.

Aneuploidy and chromosomal instability are both commonly found in cancer. Chromosomal instability leads to karyotype heterogeneity in tumors and is associated with therapy resistance, metastasis and poor prognosis. It has been hypothesized that aneuploidy per se is sufficient to drive CIN, however due to limited models and heterogenous results, it has remained controversial which aspects of aneuploidy can drive CIN. In this study we systematically tested the impact of different types of aneuploidies on the induction of CIN. We generated a plethora of isogenic aneuploid clones harboring whole chromosome or segmental aneuploidies in human p53-deficient RPE-1 cells. We observed increased segregation errors in cells harboring trisomies that strongly correlated to the number of gained genes. Strikingly, we found that clones harboring only monosomies do not induce a CIN phenotype. Finally, we found that an initial chromosome breakage event and subsequent fusion can instigate breakage-fusion-bridge cycles. By investigating the impact of monosomies, trisomies and segmental aneuploidies on chromosomal instability we further deciphered the complex relationship between aneuploidy and CIN.

DOI: https://doi.org/10.1371/journal.pone.0268579