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Centromere protein

Jackie Lang, Adrienne Barber, Sue Biggins
Chromosome segregation depends on the kinetochore, the machine that establishes force-bearing attachments between DNA and spindle microtubules. Kinetochores are formed every cell cycle via a highly regulated process that requires coordinated assembly of multiple subcomplexes on specialized chromatin. To elucidate the underlying mechanisms, we developed an assay to assemble kinetochores de novo using centromeric DNA and budding yeast extracts. Assembly is enhanced by mitotic phosphorylation of the Dsn1 kinetochore protein and generates kinetochores capable of binding microtubules...
August 17, 2018: ELife
Kohta Yoshida, Asano Ishikawa, Atsushi Toyoda, Shuji Shigenobu, Asao Fujiyama, Jun Kitano
Intragenomic conflict, the conflict of interest between different genomic regions within an individual, is proposed as a mechanism driving both the rapid evolution of heterochromatin-related proteins and the establishment of intrinsic genomic incompatibility between species. Although molecular studies of laboratory model organisms have demonstrated the link between heterochromatin evolution and hybrid abnormalities, we know little about their link in natural systems. Previously, we showed that F1 hybrids between the Japan Sea stickleback and the Pacific Ocean stickleback show hybrid male sterility and found a region responsible for hybrid male sterility on the X chromosome, but did not identify any candidate genes...
August 17, 2018: Molecular Ecology
Julie K Monda, Iain M Cheeseman
Accurate chromosome segregation critically depends on the formation of attachments between microtubule polymers and each sister chromatid. The kinetochore is the macromolecular complex that assembles at the centromere of each chromosome during mitosis and serves as the link between the DNA and the microtubules. In this Cell Science at a Glance article and accompanying poster, we discuss the activities and molecular players that are involved in generating kinetochore-microtubule attachments, including the initial stages of lateral kinetochore-microtubule interactions and maturation to stabilized end-on attachments...
August 16, 2018: Journal of Cell Science
Gabriel Keeble-Gagnère, Philippe Rigault, Josquin Tibbits, Raj Pasam, Matthew Hayden, Kerrie Forrest, Zeev Frenkel, Abraham Korol, B Emma Huang, Colin Cavanagh, Jen Taylor, Michael Abrouk, Andrew Sharpe, David Konkin, Pierre Sourdille, Benoît Darrier, Frédéric Choulet, Aurélien Bernard, Simone Rochfort, Adam Dimech, Nathan Watson-Haigh, Ute Baumann, Paul Eckermann, Delphine Fleury, Angela Juhasz, Sébastien Boisvert, Marc-Alexandre Nolin, Jaroslav Doležel, Hana Šimková, Helena Toegelová, Jan Šafář, Ming-Cheng Luo, Francisco Câmara, Matthias Pfeifer, Don Isdale, Johan Nyström-Persson, Iwgsc, Dal-Hoe Koo, Matthew Tinning, Dangqun Cui, Zhengang Ru, Rudi Appels
BACKGROUND: Numerous scaffold-level sequences for wheat are now being released and, in this context, we report on a strategy for improving the overall assembly to a level comparable to that of the human genome. RESULTS: Using chromosome 7A of wheat as a model, sequence-finished megabase-scale sections of this chromosome were established by combining a new independent assembly using a bacterial artificial chromosome (BAC)-based physical map, BAC pool paired-end sequencing, chromosome-arm-specific mate-pair sequencing and Bionano optical mapping with the International Wheat Genome Sequencing Consortium RefSeq v1...
August 17, 2018: Genome Biology
Shin-Ichiro Hiraga, Chandre Monerawela, Yuki Katou, Sophie Shaw, Kate Rm Clark, Katsuhiko Shirahige, Anne D Donaldson
Despite its evolutionarily conserved function in controlling DNA replication, the chromosomal binding sites of the budding yeast Rif1 protein are not well understood. Here, we analyse genome-wide binding of budding yeast Rif1 by chromatin immunoprecipitation, during G1 phase and in S phase with replication progressing normally or blocked by hydroxyurea. Rif1 associates strongly with telomeres through interaction with Rap1. By comparing genomic binding of wild-type Rif1 and truncated Rif1 lacking the Rap1-interaction domain, we identify hundreds of Rap1-dependent and Rap1-independent chromosome interaction sites...
August 13, 2018: EMBO Reports
Shinsuke Aida, Junko Aida, Miho Naoi, Mai Kato, Yukio Tsuura, Ichiro Natsume, Kaiyo Takubo
We estimated the telomere lengths of neoplastic and non-neoplastic mesothelial cells and examined their correlation with asbestos exposure and the expression of markers of mesothelial malignancy. Cell blocks of pleural effusion obtained from 35 cases of non-neoplastic disease (NN), 12 cases of malignant mesothelioma (MM) and 12 cases of carcinomatous effusion due to lung adenocarcinoma (LA) were examined. Fifteen of the 35 NN cases had pleural plaques (NNpp+) suggestive of asbestos exposure, and the other 20 cases had no pleural plaques (NNpp-)...
August 10, 2018: Pathology International
So Maezawa, Kazuteru Hasegawa, Kris G Alavattam, Mayuka Funakoshi, Taiga Sato, Artem Barski, Satoshi H Namekawa
Spermatogenesis involves the progressive reorganization of heterochromatin. However, the mechanisms that underlie the dynamic remodeling of heterochromatin remain unknown. Here, we identify SCML2, a germline-specific Polycomb protein, as a critical regulator of heterochromatin organization in spermatogenesis. We show that SCML2 accumulates on pericentromeric heterochromatin (PCH) in male germ cells, where it suppresses PRC1-mediated monoubiquitination of histone H2A at Lysine 119 (H2AK119ub) and promotes deposition of PRC2-mediated H3K27me3 during meiosis...
August 10, 2018: Journal of Cell Science
Emily L Kurdzo, Hoa H Chuong, Jared M Evatt, Dean S Dawson
In meiosis I, homologous chromosomes segregate away from each other-the first of two rounds of chromosome segregation that allow the formation of haploid gametes. In prophase I, homologous partners become joined along their length by the synaptonemal complex (SC) and crossovers form between the homologs to generate links called chiasmata. The chiasmata allow the homologs to act as a single unit, called a bivalent, as the chromosomes attach to the microtubules that will ultimately pull them away from each other at anaphase I...
August 9, 2018: PLoS Genetics
Alexey K Mazur, Eugene Gladyshev
In some fungi, a premeiotic process known as repeat-induced point mutation (RIP) can accurately identify and mutate nearly all gene-sized DNA repeats present in the haploid germline nuclei. Studies in Neurospora crassa have suggested that RIP detects sequence homology directly between intact DNA double helices, without strand separation and without the participation of RecA-like proteins. Those studies used the aggregated number of RIP mutations as a simple quantitative measure of RIP activity. Additional structural information about homologous DNA-DNA pairing during RIP can be extracted by analyzing spatial distributions of RIP mutations converted into profiles of partitioned RIP propensity (PRP)...
July 21, 2018: Biophysical Journal
Joyce J Thompson, Rupinder Kaur, Carlos P Sosa, Jeong-Heon Lee, Katsunobu Kashiwagi, Dan Zhou, Keith D Robertson
The interplay between transcription factors and epigenetic writers like the DNA methyltransferases (DNMTs), and the role of this interplay in gene expression, is being increasingly appreciated. ZBTB24, a poorly characterized zinc-finger protein, or the de novo methyltransferase DNMT3B, when mutated, cause Immunodeficiency, Centromere Instability, and Facial anomalies (ICF) syndrome, suggesting an underlying mechanistic link. Chromatin immunoprecipitation coupled with loss-of-function approaches in model systems revealed common loci bound by ZBTB24 and DNMT3B, where they function to regulate gene body methylation...
July 31, 2018: Nucleic Acids Research
Joan M Martínez-Láinez, David F Moreno, Eva Parisi, Josep Clotet, Martí Aldea
Cell size scales with ploidy in a great range of eukaryotes, but the underlying mechanisms remain unknown. Using various orthogonal single-cell approaches, we show that cell size increases linearly with centromere (CEN) copy number in budding yeast. This effect is due to a G1 delay mediated by increased degradation of Cln3, the most upstream G1 cyclin acting at Start, and specific centromeric signaling proteins, namely Mad3 and Bub3. Mad3 binds both Cln3 and Cdc4, the adaptor component of the Skp1/Cul1/F-box (SCF) complex that targets Cln3 for degradation, these interactions being essential for the CEN-dosage dependent effects on cell size...
August 6, 2018: PLoS Biology
Marco Novais-Cruz, Maria Alba Abad, Wilfred Fj van IJcken, Niels Galjart, A Arockia Jeyaprakash, Helder Maiato, Cristina Ferrás
Recent studies have challenged the prevailing dogma that transcription is repressed during mitosis. Transcription was also proposed to sustain a robust spindle assembly checkpoint (SAC) response. Here we used live-cell imaging of human cells, RNA-seq and qPCR to investigate the requirement for de novo transcription during mitosis. Under conditions of persistently unattached kinetochores, transcription inhibition with actinomycin D, or treatment with other DNA-intercalating drugs, delocalized the chromosomal passenger complex (CPC) protein Aurora B from centromeres, compromising SAC signalling and cell fate...
August 6, 2018: ELife
Mridula Nambiar, Gerald R Smith
In most eukaryotes, meiotic crossovers are essential for error-free chromosome segregation but are specifically repressed near centromeres to prevent missegregation. Recognized for >85 years, the molecular mechanism of this repression has remained unknown. Meiotic chromosomes contain two distinct cohesin complexes: pericentric complex (for segregation) and chromosomal arm complex (for crossing over). We show that the pericentric-specific complex also actively represses pericentric meiotic double-strand break (DSB) formation and, consequently, crossovers...
August 16, 2018: Molecular Cell
Natalay Kouprina, Nikolai Petrov, Oscar Molina, Mikhail Liskovykh, Elisa Pesenti, Jun-Ichirou Ohzeki, Hiroshi Masumoto, William C Earnshaw, Vladimir Larionov
Since their description in the late 1990s, Human Artificial Chromosomes (HACs) bearing functional kinetochores have been considered as promising systems for gene delivery and expression. More recently a HAC assembled from a synthetic alphoid DNA array has been exploited in studies of centromeric chromatin and in assessing the impact of different epigenetic modifications on kinetochore structure and function in human cells. This HAC was termed the alphoidtetO -HAC, as the synthetic monomers each contained a tetO sequence in place of the CENP-B box that can be targeted specifically with tetR-fusion proteins...
August 16, 2018: ACS Synthetic Biology
Sebastian Hoffmann, Daniele Fachinetti
Measuring protein dynamics is essential to uncover protein function and to understand the formation of large protein complexes such as centromeres. Recently, genome engineering in human cells has improved our ability to study the function of endogenous proteins. By combining genome editing techniques with the auxin-inducible degradation (AID) system, we created a versatile tool to study protein dynamics. This system allows us to analyze both protein function and dynamics by enabling rapid protein depletion and reexpression in the same experimental setup...
2018: Methods in Molecular Biology
Guoliang Yuan, Behzad Heidari, Shinichiro Komaki, Arp Schnittger, Cathrine Lillo, Nico De Storme, Danny N V Geelen
Correct separation of homologous chromosomes during meiosis I and sister chromatids during meiosis II relies on the tight control of the cohesion complex that holds the sister chromatids together. Phosphorylation and subsequent cleavage of meiotic recombination protein REC8 (REC8-like family protein [SYN1] in Arabidopsis thaliana), the alpha kleisin subunit of the cohesion ring, along the chromosome arms at meiosis I allows crossovers (COs) and separation of homologous chromosomes without chromatid dissociation...
July 30, 2018: Plant Physiology
Amr Ahmed El-Arabey, Salama Abdu Salama, Adel Rashad Abd-Allah
In 2015, more than 1.6 million new cancer cases with 589,430 deaths were estimated over worldwide. Cancer is a complex disease with abnormal cell growth control which is hallmarked by chromosome misalignment and consequently genomic instability. Mitosis is a well-known target for chemotherapy as taxol and colchicines inhibit tumor cell division by inhibiting mitotic spindle plasticity. Accumulating evidence has revealed that the Centromere-associated protein E (CENP-E) is expressed during mitosis and plays critical roles in inaccurate chromosome alignment...
July 19, 2018: Life Sciences
Yuting Chen, Na Zhu, Yuyi Luo, Keqi Hu, Yungang Liu
Polychlorinated biphenyls (PCBs) as a group of persistent organic pollutants are confirmed human carcinogens; however, their mutagenicity remains mostly unknown. We have reported the mutagenicity of some PCBs with one to four chlorines in mammalian cells expressing human CYP2E1. To further explore the structural requirements for the mutagenicity of PCBs, eight tri- and tetrachlorobiphenyls untested before were investigated for the induction of gene mutations and micronuclei in a V79-derived cell line expressing both human CYP2E1 and sulfotransferase (SULT) 1A1 (V79-hCYP2E1-hSULT1A1), with SULT1A1 activity inhibited by pentachlorophenol, a potent SULT1 inhibitor...
July 11, 2018: Chemosphere
Hongda Chen, Yue Zhang, Siwen Li, Ni Li, Yuhan Chen, Bei Zhang, Chunfeng Qu, Huiguo Ding, Jian Huang, Min Dai
Background: Although a number of serum biomarkers for detection of hepatocellular carcinoma (HCC) have been explored, their exact diagnostic value remains unclear. We aimed to conduct a direct comparison of five representative serum biomarkers for detecting HCC and to derive multi-marker prediction algorithms. Patients and methods: In total, 846 patients were recruited from three hospitals in China, including 202 HCC patients, 226 liver cirrhosis patients, 215 chronic hepatitis B virus-infected patients, and 203 healthy volunteers...
2018: Cancer Management and Research
Saima Akram, Fengrui Yang, Junying Li, Gregory Adams, Yingying Liu, Xiaoxuan Zhuang, Lingluo Chu, Xu Liu, Nerimah Emmett, Winston Thompson, McKay Mullen, Saravana Muthusamy, Wenwen Wang, Fei Mo, Xing Liu
Heterochromatin protein 1α (HP1α) regulates chromatin specification and plasticity during cell fate decision. Different structural determinants account for HP1α localization and function during cell division cycle. Our earlier study showed that centromeric localization of HP1α depends on the epigenetic mark H3K9me3 in interphase, while its centromeric location in mitosis relies on uncharacterized PXVXL-containing factors. Here, we identified a PXVXL-containing protein, ligand-dependent nuclear receptor-interacting factor 1 (LRIF1), which recruits HP1α to the centromere of mitotic chromosomes and its interaction with HP1α is essential for accurate chromosome segregation during mitosis...
July 16, 2018: Journal of Molecular Cell Biology
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