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Spindle checkpoint

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https://www.readbyqxmd.com/read/29321280/analysis-of-mitotic-checkpoint-function-in-xenopus-egg-extracts
#1
Yinghui Mao
Accurate sister chromatid segregation is pivotal in the faithful transmission of genetic information during each cell division. To ensure accurate segregation, eukaryotic organisms have evolved a "mitotic (or spindle assembly) checkpoint" to prevent premature advance to anaphase before successful attachment of every chromosome to the microtubules of the mitotic spindle. An unattached kinetochore generates a diffusible signal that inhibits ubiquitination of substrates such as cyclin B and securins. This protocol presents an in vitro assay for studying the mitotic checkpoint using Xenopus laevis egg extracts...
January 10, 2018: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/29285250/spc24-promotes-osteosarcoma-progression-by-increasing-egfr-mapk-signaling
#2
Jun Sheng, Mengchen Yin, Zhengwang Sun, Xia Kang, Da Liu, Kai Jiang, Jia Xu, Feixing Zhao, Qunfeng Guo, Wei Zheng
In this study, we investigated the role of the spindle checkpoint protein SPC24 in osteosarcoma progression. SPC24 knockdown in 143B and U2OS osteosarcoma cells decreased cell growth, survival and invasiveness. The SPC24 knockdown cells also exhibited low EGFR, Ras and phospho-ERK levels and high E-cadherin levels, suggesting inhibition of EGFR/Ras/ERK signaling and epithelial-to-mesenchymal transitioning. Xenografted SPC24 knockdown osteosarcoma cells showed reduced tumor growth in nude mice with decreased EGFR and phospho-ERK levels and increased E-cadherin levels...
December 1, 2017: Oncotarget
https://www.readbyqxmd.com/read/29283376/differential-selective-pressures-experienced-by-the-aurora-kinase-gene-family
#3
Joni M Seeling, Alexis A Farmer, Adam Mansfield, Hyuk Cho, Madhusudan Choudhary
Aurora kinases (AKs) are serine/threonine kinases that are essential for cell division. Humans have three AK genes: AKA, AKB, and AKC. AKA is required for centrosome assembly, centrosome separation, and bipolar spindle assembly, and its mutation leads to abnormal spindle morphology. AKB is required for the spindle checkpoint and proper cytokinesis, and mutations cause chromosome misalignment and cytokinesis failure. AKC is expressed in germ cells, and has a role in meiosis analogous to that of AKB in mitosis...
December 28, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29276128/tension-induced-error-correction-and-not-kinetochore-attachment-status-activates-the-sac-in-an-aurora-b-c-dependent-manner-in-oocytes
#4
Antoine Vallot, Ioanna Leontiou, Damien Cladière, Warif El Yakoubi, Susanne Bolte, Eulalie Buffin, Katja Wassmann
Cell division with partitioning of the genetic material should take place only when paired chromosomes named bivalents (meiosis I) or sister chromatids (mitosis and meiosis II) are correctly attached to the bipolar spindle in a tension-generating manner. For this to happen, the spindle assembly checkpoint (SAC) checks whether unattached kinetochores are present, in which case anaphase onset is delayed to permit further establishment of attachments. Additionally, microtubules are stabilized when they are attached and under tension...
December 16, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/29244163/regulation-of-chromosome-segregation-in-oocytes-and-the-cellular-basis-for-female-meiotic-errors
#5
Jessica Greaney, Zhe Wei, Hayden Homer
BACKGROUND: Meiotic chromosome segregation in human oocytes is notoriously error-prone, especially with ageing. Such errors markedly reduce the reproductive chances of increasing numbers of women embarking on pregnancy later in life. However, understanding the basis for these errors is hampered by limited access to human oocytes. OBJECTIVE AND RATIONALE: Important new discoveries have arisen from molecular analyses of human female recombination and aneuploidy along with high-resolution analyses of human oocyte maturation and mouse models...
December 13, 2017: Human Reproduction Update
https://www.readbyqxmd.com/read/29234286/identification-of-biomarkers-correlated-with-the-tnm-staging-and-overall-survival-of-patients-with-bladder-cancer
#6
Sheng Li, Xiaoping Liu, Tongzu Liu, Xiangyu Meng, Xiaohong Yin, Cheng Fang, Di Huang, Yue Cao, Hong Weng, Xiantao Zeng, Xinghuan Wang
Objective: To identify candidate biomarkers correlated with clinical prognosis of patients with bladder cancer (BC). Methods: Weighted gene co-expression network analysis was applied to build a co-expression network to identify hub genes correlated with tumor node metastasis (TNM) staging of BC patients. Functional enrichment analysis was conducted to functionally annotate the hub genes. Protein-protein interaction network analysis of hub genes was performed to identify the interactions among the hub genes...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/29228672/pten-regulates-spindle-assembly-checkpoint-timing-through-mad1-in-interphase
#7
Yu Liu, Xiao Du, Shuting Zhang, Yang Liu, Qiaoling Zhang, Qi Yin, Michael A McNutt, Yuxin Yin
The spindle assembly checkpoint (SAC) restrains anaphase progression to ensure all chromosomes attach properly to the spindle. Although SAC timing has been extensively investigated in mitosis, its mechanism of regulation in interphase is unclear. We report that PTEN functions as a crucial activator of SAC timing and protects chromosome segregation under both spindle poison treated and untreated conditions. We show that PTEN physically interacts with MAD1 and promotes its dimerization and localization in the nuclear pore...
November 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/29228610/targeting-polo-like-kinase-1-in-smarcb1-deleted-atypical-teratoid-rhabdoid-tumor
#8
Irina Alimova, Angela M Pierce, Peter Harris, Andrew Donson, Diane K Birks, Eric Prince, Ilango Balakrishnan, Nicholas K Foreman, Marcel Kool, Lindsey Hoffman, Sujatha Venkataraman, Rajeev Vibhakar
Atypical teratoid rhabdoid tumor (ATRT) is an aggressive and malignant pediatric brain tumor. Polo-like kinase 1 (PLK1) is highly expressed in many cancers and essential for mitosis. Overexpression of PLK1 promotes chromosome instability and aneuploidy by overriding the G2-M DNA damage and spindle checkpoints. Recent studies suggest that targeting PLK1 by small molecule inhibitors is a promising approach to tumor therapy. We investigated the effect of PLK1 inhibition in ATRT. Gene expression analysis showed that PLK1 was overexpressed in ATRT patient samples and tumor cell lines...
November 14, 2017: Oncotarget
https://www.readbyqxmd.com/read/29220700/prognostic-importance-of-aurora-kinases-and-mitotic-spindle-genes-transcript-levels-in-myelodysplastic-syndrome
#9
Daniela de Paula Borges, Antônio Wesley Araújo Dos Santos, Carlos Roberto Koscky Paier, Howard Lopes Ribeiro, Marília Braga Costa, Izabelle Rocha Farias, Roberta Taiane Germano de Oliveira, Ivo Gabriel da Frota França, Gabrielle Melo Cavalcante, Sílvia Maria Meira Magalhães, Ronald Feitosa Pinheiro
Myelodysplastic syndrome (MDS) are a heterogeneous group of clonal disease characterized by insufficiency of bone marrow, increase of apoptosis and increased risk of acute leukemia progression. Proteins related to the mitotic spindle (AURKA, AURKB, TPX2), to the mitotic checkpoint (MAD2, CDC20) and the regulation of the cell cycle (p21) are directly related to chromosomal stability and tumor development. This study aimed to evaluate the mRNA expression levels of these genes in 101 MDS patients using a real-time PCR methodology...
November 28, 2017: Leukemia Research
https://www.readbyqxmd.com/read/29209640/some-assembly-required-redefining-the-mitotic-checkpoint
#10
John C Meadows, Jonathan B A Millar
The spindle assembly checkpoint (also known as the spindle or mitotic checkpoint) is a surveillance system that ensures fidelity of chromosome segregation. Here we suggest, in light of historical and more recent evidence, that this signaling system monitors kinetochore attachment and spindle assembly by two distinct, but functionally overlapping, pathways.
2017: Molecular & Cellular Oncology
https://www.readbyqxmd.com/read/29208896/mechanistic-insight-into-trip13-catalyzed-mad2-structural-transition-and-spindle-checkpoint-silencing
#11
Melissa L Brulotte, Byung-Cheon Jeong, Faxiang Li, Bing Li, Eric B Yu, Qiong Wu, Chad A Brautigam, Hongtao Yu, Xuelian Luo
The spindle checkpoint maintains genomic stability and prevents aneuploidy. Unattached kinetochores convert the latent open conformer of the checkpoint protein Mad2 (O-Mad2) to the active closed conformer (C-Mad2), bound to Cdc20. C-Mad2-Cdc20 is incorporated into the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex/cyclosome (APC/C). The C-Mad2-binding protein p31comet and the ATPase TRIP13 promote MCC disassembly and checkpoint silencing. Here, using nuclear magnetic resonance (NMR) spectroscopy, we show that TRIP13 and p31comet catalyze the conversion of C-Mad2 to O-Mad2, without disrupting its stably folded core...
December 5, 2017: Nature Communications
https://www.readbyqxmd.com/read/29196757/mitotic-slippage-and-the-subsequent-cell-fates-after-inhibition-of-aurora-b-during-tubulin-binding-agent-induced-mitotic-arrest
#12
Yasuo Tsuda, Makoto Iimori, Yuichiro Nakashima, Ryota Nakanishi, Koji Ando, Kippei Ohgaki, Hiroyuki Kitao, Hiroshi Saeki, Eiji Oki, Yoshihiko Maehara
Tubulin-binding agents (TBAs) are designed to target microtubule (MT) dynamics, resulting in compromised mitotic spindles and an unsatisfied spindle assembly checkpoint. The activity of Aurora B kinase is indispensable for TBA-induced mitotic arrest, and its inhibition causes mitotic slippage and postmitotic endoreduplication. However, the precise phenomenon underlying mitotic slippage, which is caused by treatment with both Aurora B inhibitors and TBAs, and the cell fate after postmitotic slippage are not completely understood...
December 1, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29196303/whole-chromosome-loss-and-associated-breakage-fusion-bridge-cycles-transform-mouse-tetraploid-cells
#13
Rozario Thomas, Daniel Henry Marks, Yvette Chin, Robert Benezra
Whole chromosome gains or losses (aneuploidy) are a hallmark of ~70% of human tumors. Modeling the consequences of aneuploidy has relied on perturbing spindle assembly checkpoint (SAC) components, but interpretations of these experiments are clouded by the multiple functions of these proteins. Here, we used a Cre recombinase-mediated chromosome loss strategy to individually delete mouse chromosomes 9, 10, 12, or 14 in tetraploid immortalized murine embryonic fibroblasts. This methodology also involves the generation of a dicentric chromosome intermediate, which subsequently undergoes a series of breakage-fusion-bridge (BFB) cycles...
December 1, 2017: EMBO Journal
https://www.readbyqxmd.com/read/29192061/nude-l-regulates-dynein-at-kinetochores-but-is-dispensable-for-other-dynein-functions-in-the-c-elegans-early-embryo
#14
Patrícia A Simões, Ricardo Celestino, Ana X Carvalho, Reto Gassmann
In mitosis, the molecular motor dynein is recruited to kinetochores by the Rod-Zw10-Zwilch complex (RZZ) and Spindly to control spindle assembly checkpoint (SAC) signaling and microtubule attachment. How the ubiquitous dynein co-factors Lis1 and NudE/L contribute to these functions remains poorly understood. Here, we show that the C. elegans NudE/L homolog NUD-2 is dispensable for dynein- and LIS-1-dependent mitotic spindle assembly in the zygote. This facilitates functional characterization of kinetochore-localized NUD-2, which is recruited by the CENP-F-like proteins HCP-1/2 independently of RZZ-Spindly and dynein-LIS-1...
November 30, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29190829/a-comprehensive-analysis-of-breast-cancer-microbiota-and-host-gene-expression
#15
Kevin J Thompson, James N Ingle, Xiaojia Tang, Nicholas Chia, Patricio R Jeraldo, Marina R Walther-Antonio, Karunya K Kandimalla, Stephen Johnson, Janet Z Yao, Sean C Harrington, Vera J Suman, Liewei Wang, Richard L Weinshilboum, Judy C Boughey, Jean-Pierre Kocher, Heidi Nelson, Matthew P Goetz, Krishna R Kalari
The inflammatory tumoral-immune response alters the physiology of the tumor microenvironment, which may attenuate genomic instability. In addition to inducing inflammatory immune responses, several pathogenic bacteria produce genotoxins. However the extent of microbial contribution to the tumor microenvironment biology remains unknown. We utilized The Cancer Genome Atlas, (TCGA) breast cancer data to perform a novel experiment utilizing unmapped and mapped RNA sequencing read evidence to minimize laboratory costs and effort...
2017: PloS One
https://www.readbyqxmd.com/read/29186573/conformational-dynamics-of-the-hop1-horma-domain-reveal-a-common-mechanism-with-the-spindle-checkpoint-protein-mad2
#16
Alan M V West, Elizabeth A Komives, Kevin D Corbett
The HORMA domain is a highly conserved protein-protein interaction module found in eukaryotic signaling proteins including the spindle assembly checkpoint protein Mad2 and the meiotic HORMAD proteins. HORMA domain proteins interact with short 'closure motifs' in partner proteins by wrapping their C-terminal 'safety belt' region entirely around these motifs, forming topologically-closed complexes. Closure motif binding and release requires large-scale conformational changes in the HORMA domain, but such changes have only been observed in Mad2...
November 25, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29180432/phosphorylation-of-cenp-c-by-aurora-b-facilitates-kinetochore-attachment-error-correction-in-mitosis
#17
Xing Zhou, Fan Zheng, Chengliang Wang, Minhao Wu, Xiaozhen Zhang, Qian Wang, Xuebiao Yao, Chuanhai Fu, Xuan Zhang, Jianye Zang
Kinetochores are superprotein complexes that orchestrate chromosome segregation via a dynamic interaction with spindle microtubules. A physical connection between CENP-C and the Mis12-Ndc80-Knl1 (KMN) protein network is an important pathway that is used to assemble kinetochores on CENP-A nucleosomes. Multiple outer kinetochore components are phosphorylated by Aurora B kinase to activate the spindle assembly checkpoint (SAC) and to ensure accurate chromosome segregation. However, it is unknown whether Aurora B can phosphorylate inner kinetochore components to facilitate proper mitotic chromosome segregation...
November 27, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29177745/monitoring-the-dna-topoisomerase-ii-checkpoint-in-saccharomyces-cerevisiae
#18
Katherine Furniss, Amit C J Vas, Andrew B Lane, Duncan J Clarke
Topoisomerase II activity is crucial to maintain genome stability through the removal of catenanes in the DNA formed during DNA replication and scaffolding the mitotic chromosome. Perturbed Topo II activity causes defects in chromosome segregation due to persistent catenations and aberrant DNA condensation during mitosis. Recently, novel top2 alleles in the yeast Saccharomyces cerevisiae revealed a checkpoint control which responds to perturbed Topo II activity. Described in this chapter are protocols for assaying the phenotypes seen in top2 mutants on a cell biological basis in live cells: activation of the Topo II checkpoint using spindle morphology, chromosome condensation using fluorescently labeled chromosomal loci and cell cycle progression by flow cytometry...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29167309/visualizing-the-complex-functions-and-mechanisms-of-the-anaphase-promoting-complex-cyclosome-apc-c
#19
REVIEW
Claudio Alfieri, Suyang Zhang, David Barford
The anaphase promoting complex or cyclosome (APC/C) is a large multi-subunit E3 ubiquitin ligase that orchestrates cell cycle progression by mediating the degradation of important cell cycle regulators. During the two decades since its discovery, much has been learnt concerning its role in recognizing and ubiquitinating specific proteins in a cell-cycle-dependent manner, the mechanisms governing substrate specificity, the catalytic process of assembling polyubiquitin chains on its target proteins, and its regulation by phosphorylation and the spindle assembly checkpoint...
November 2017: Open Biology
https://www.readbyqxmd.com/read/29165592/folate-deficiency-induces-mitotic-aberrations-and-chromosomal-instability-by-compromising-the-spindle-assembly-checkpoint-in-cultured-human-colon-cells
#20
Xihan Guo, Juan Ni, Yuqian Zhu, Tao Zhou, Xiaoling Ma, Jinglun Xue, Xu Wang
Folates comprise the essential B9 vitamin that act as cofactors and cosubstrates in one-carbon metabolism for both biosynthesis and methylation of DNA and RNA. Folate deficiency (FD) has been shown to induce chromosomal instability (CIN), yet the underlying mechanisms are poorly understood. Here, we used human NCM460 colon mucosal cells as a model to investigate the effect of FD on spindle assembly checkpoint (SAC), a cell-cycle regulatory pathway preventing CIN during mitosis. Cells were maintained in medium containing 1...
November 19, 2017: Mutagenesis
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