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Anna A Ye, Stuart Cane, Thomas J Maresca
High-fidelity transmission of the genome through cell division requires that all sister kinetochores bind to dynamic microtubules (MTs) from opposite spindle poles. The application of opposing forces to this bioriented configuration produces tension that stabilizes kinetochore-microtubule (kt-MT) attachments. Defining the magnitude of force that is applied to kinetochores is central to understanding the mechano-molecular underpinnings of chromosome segregation; however, existing kinetochore force measurements span orders of magnitude...
October 20, 2016: Nature Communications
Britta Unruhe-Knauf, Shirley K Knauer
The chromosomal passenger complex (CPC) is a key regulator of cell division. Its proper localization during the different phases of mitosis and cytokinesis is crucial for the exertion of its various functions. HDACi treatment has been demonstrated to disturb the centromeric localization of the CPC in tumor cells, thus leading to severe mitotic defects often followed by apoptosis. In this chapter, we describe how HDACi-induced changes of the CPC localization can be analyzed by indirect immunofluorescence using CPC-specific primary and fluorophore-coupled secondary antibodies followed by confocal microscopy...
2017: Methods in Molecular Biology
Emilie Lukasova, Aleš Kovařík, Alena Bačíková, Martin Falk, Stanislav Kozubek
The cellular transition to senescence is associated with extensive chromatin reorganization and gene expression changes. Recent studies appeared implying an association of lamin B1 (LB1) reduction with chromatin rearrangement in human fibroblasts promoted to senescence, while the mechanisms and structural features of these relations were not yet clarified. In this work we examined the functions of LB1 and lamin B receptor (LBR) in human cancer cells. We found that both LB1 and LBR tend to deplete during cancer cells transfer to senescence by γ-irradiation...
October 19, 2016: Biochemical Journal
Olga I Podgornaya, Irina N Vasilyeva, Vladimir G Bespalov
Only limited sequencing data of the normal extracellular DNA (ecDNA) are currently available. The uptake of the ecDNA by cultured cells and its integration into the host chromatin have been demonstrated. A number of membrane-bearing vesicles in plasma and serum have been shown to carry nucleic acids. The presence of Tandem Repeat (TR) in both apoptotic DNA of HUVEC culture medium and membrane-associated DNA is shown. The existence and successful application of CREST serum also show the presence of fragments of the centromeric heterochromatin together with their TR and specific proteins in blood...
2016: Advances in Experimental Medicine and Biology
J-B Monfort, I Lazareth, P Priollet
OBJECTIVE: Association between cancer and systemic sclerosis (SSc) has been described. However, paraneoplastic SSc is not well known. The aim of this article is to describe cases of paraneoplastic systemic sclerosis and to compare them to other cases of the literature, to find characteristics that can make suspect a paraneoplastic mechanism when SSc is diagnosed. METHODS: We retrospectively analyzed patients, in our department who, over the last 15 years, presented with Raynaud's phenomenon with a diagnosis of SSc (including cancer during the period of SSc)...
October 14, 2016: Journal des Maladies Vasculaires
Cecilia Andreu, Marcel Lí Del Olmo
Yeast surface display is a powerful tool widely used for many biotechnological and biomedical applications. It consists in exposing peptides and proteins of interest on the surface of Saccharomyces cerevisiae and other yeasts. These molecules are fused to the amino or carboxy terminus of an appropriate cell wall protein, usually bound by glycosylphosphatidylinositol. Several systems for this purpose have been reported to date. In this work, we describe a new yeast surface display strategy based on cell wall protein Spi1 as an anchor, which is expressed in centromeric and episomal plasmids under the control of its own promoter or that corresponding to the PGK1 glycolytic gene...
October 15, 2016: Applied Microbiology and Biotechnology
Paul Knopp, Yvonne D Krom, Christopher R S Banerji, Maryna Panamarova, Louise A Moyle, Bianca den Hamer, Silvère M van der Maarel, Peter S Zammit
Skeletal muscle wasting in facioscapulohumeral muscular dystrophy (FSHD) results in substantial morbidity. On a disease-permissive chromosome 4qA haplotype, genomic and/or epigenetic changes at the D4Z4 macrosatellite repeat allows transcription of the DUX4 retrogene. Analysing transgenic mice carrying a human D4Z4 genomic locus from an FSHD-affected individual showed that DUX4 was transiently induced in myoblasts during skeletal muscle regeneration. Centromeric to the D4Z4 repeats is an inverted D4Z4 unit encoding DUX4c...
October 15, 2016: Journal of Cell Science
Shane C Quinonez, Thomas D Gelehrter, Wendy R Uhlmann
Small supernumerary marker chromosomes (sSMC) are abnormal chromosomes that cannot be characterized by standard banding cytogenetic techniques. A minority of sSMC contain a neocentromere, which is an ectopic centromere lacking the characteristic alpha-satellite DNA. The phenotypic manifestations of sSMC and neocentromeric sSMC are variable and range from severe intellectual disability and multiple congenital anomalies to a normal phenotype. Here we report a patient with a diagnosis of Marfan syndrome and infertility found to have an abnormal karyotype consisting of a chromosome 15 deletion and a ring-type sSMC likely stabilized by a neocentromere derived via a mechanism initially described by Barbara McClintock in 1938...
October 14, 2016: American Journal of Medical Genetics. Part A
B Roche, B Arcangioli, R A Martienssen
Quiescent cells play a predominant role in most organisms. Here, we identify RNA interference (RNAi) as a major requirement for quiescence (G0) in Schizosaccharomyces pombe RNAi mutants lose viability at G0-entry and are unable to maintain long-term quiescence. We obtained dcr1Δ G0 suppressors, which mapped to genes involved in chromosome segregation, RNA polymerase-associated factors, and heterochromatin formation. We propose a model in which RNAi promotes RNA polymerase release in cycling and quiescent cells: (i) RNA pol II release mediates heterochromatin formation at centromeres allowing proper chromosome segregation during mitotic growth and G0-entry, and (ii) RNA pol I release prevents heterochromatin formation at rDNA during quiescence maintenance...
October 13, 2016: Science
Marinela Dukić, Daniel Berner, Marius Roesti, Christoph R Haag, Dieter Ebert
BACKGROUND: Recombination rate is an essential parameter for many genetic analyses. Recombination rates are highly variable across species, populations, individuals and different genomic regions. Due to the profound influence that recombination can have on intraspecific diversity and interspecific divergence, characterization of recombination rate variation emerges as a key resource for population genomic studies and emphasises the importance of high-density genetic maps as tools for studying genome biology...
October 13, 2016: BMC Genetics
Andrew W Grenfell, Magdalena Strzelecka, Rebecca Heald
No abstract text is available yet for this article.
October 13, 2016: Cell Cycle
Erez Rechavi, Atar Lev, Eran Eyal, Ortal Barel, Nitzan Kol, Sarit Farage Barhom, Ben Pode-Shakked, Yair Anikster, Raz Somech, Amos J Simon
PURPOSE: Immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome is an extremely rare autosomal recessive disease. The immune phenotype is characterized by hypogammaglobulinemia in the presence of B cells. T cell lymphopenia also develops in some patients. We sought to further investigate the immune defect in an ICF patient with a novel missense mutation in DNMT3B and a severe phenotype. METHODS: Patient lymphocytes were examined for subset counts, immunoglobulin levels, T and B cell de novo production (via excision circles) and receptor repertoire diversity...
October 12, 2016: Journal of Clinical Immunology
Mai Takagi, Takuya Sakamoto, Ritsuko Suzuki, Keiichirou Nemoto, Takeshi Obayashi, Takeshi Hirakawa, Tomoko M Matsunaga, Daisuke Kurihara, Yuko Nariai, Takeshi Urano, Tatsuya Sawasaki, Sachihiro Matsunaga
Aurora kinase (AUR) is a well-known mitotic serine/threonine kinase that regulates centromere formation, chromosome segregation, and cytokinesis in eukaryotes. In addition to regulating mitotic events, AUR has been shown to regulate protein dynamics during interphase in animal cells. In contrast, there has been no identification and characterization of substrates and/or interacting proteins during interphase in plants. The Arabidopsis thaliana genome encodes three AUR paralogues, AtAUR1, AtAUR2, and AtAUR3...
October 12, 2016: Journal of Plant Research
Tsung-Han S Hsieh, Geoffrey Fudenberg, Anton Goloborodko, Oliver J Rando
We present Micro-C XL, an improved method for analysis of chromosome folding at mononucleosome resolution. Using long crosslinkers and isolation of insoluble chromatin, Micro-C XL increases signal-to-noise ratio. Micro-C XL maps of budding and fission yeast genomes capture both short-range chromosome fiber features such as chromosomally interacting domains and higher order features such as centromere clustering. Micro-C XL provides a single assay to interrogate chromosome folding at length scales from the nucleosome to the full genome...
October 10, 2016: Nature Methods
Petr Ráb, Cassia F Yano, Sébastien Lavoué, Oladele I Jegede, Luiz A C Bertollo, Tariq Ezaz, Zuzana Majtánová, Ezequiel A de Oliveira, Marcelo B Cioffi
The monophyletic order Osteoglossiformes represents one of the most ancestral groups of teleosts and has at least 1 representative in all continents of the southern hemisphere, with the exception of Antarctica. However, despite its phylogenetic and biogeographical importance, cytogenetic data in Osteoglossiformes are scarce. Here, karyotype and chromosomal characteristics of the lower Niger River population of the African butterfly fish Pantodon buchholzi, the sole species of the family Pantodontidae (Osteoglossiformes), were examined using conventional and molecular cytogenetic approaches...
October 7, 2016: Cytogenetic and Genome Research
R C R Noronha, L M R Barros, R E F Araújo, D F Marques, C Y Nagamachi, C Martins, J C Pieczarka
BACKGROUND: Cytogenetic studies were conducted in the Brazilian Amazon turtles, Podocnemis expansa Schweigger, 1912 (PEX) and Podocnemis unifilis Troschel, 1848 (PUN) to understand their karyoevolution. Their chromosomal complements were compared using banding techniques (C, G-, Ag-NOR and Chromomycin A3) and fluorescence in situ hybridization (FISH), and efforts were made to establish evolutionary chromosomal relationships within the Podocnemidae family. RESULTS: Our results revealed that both species have a chromosome complement of 2n = 28...
2016: Molecular Cytogenetics
Qinbo Qin, Zhengfa Lai, Liu Cao, Qiong Xiao, YuDe Wang, Shaojun Liu
To better understand genomic changes in the early generations after polyploidisation, we examined the chromosomal consequences of genomic merger in allotetraploid hybrids (4 nF1) (AABB, 4n = 148) of Carassius auratus red var. (RCC) (AA, 2n = 100) (♀) × Megalobrama amblycephala (BSB) (BB, 2n = 48) (♂). Complete loss of the paternal 5S rDNA sequence and the expected number of maternal chromosomal loci were found in 4 nF1, suggesting directional genomic changes occurred in the first generations after polyploidisation...
October 5, 2016: Scientific Reports
Yijun Zhang, Youguang Luo, Rui Lyu, Jie Chen, Ruming Liu, Dengwen Li, Min Liu, Jun Zhou
Cell migration, a complex process critical for tumor progression and metastasis, requires a dynamic crosstalk between microtubules (MTs) and focal adhesions (FAs). However, the molecular mechanisms underlying this event remain elusive. Herein we identify the proto-oncogenic protein Src as an important player in the regulation of the MT-FA crosstalk. Src interacts with and phosphorylates end-binding protein 1 (EB1), a member of MT plus end-tracking proteins (+TIPs), both in cells and in vitro. Systematic mutagenesis reveals that tyrosine-247 (Y247) is the primary residue of EB1 phosphorylated by Src...
2016: Theranostics
Hugh P Cam, Simon Whitehall
Reporter gene silencing assays provide a facile method for assessing the function of heterochromatin in Schizosaccharomyces pombe They use strains containing auxotrophic markers (commonly ura4(+) or ade6(+)) located within a heterochromatic region. Transcriptional silencing of these reporters can be assessed by plating serial dilutions of cells onto minimal agar. In addition, silencing of ura4(+) renders cells resistant to 5-fluoroorotic acid (5-FOA) and ade6(+) silencing results in red colony color on adenine-limiting agar...
October 3, 2016: Cold Spring Harbor Protocols
Patrick M Redli, Ivana Gasic, Patrick Meraldi, Erich A Nigg, Anna Santamaria
Chromosome biorientation and accurate segregation rely on the plasticity of kinetochore-microtubule (KT-MT) attachments. Aurora B facilitates KT-MT dynamics by phosphorylating kinetochore proteins that are critical for KT-MT interactions. Among the substrates whose microtubule and kinetochore binding is curtailed by Aurora B is the spindle and kinetochore-associated (Ska) complex, a key factor for KT-MT stability. Here, we show that Ska is not only a substrate of Aurora B, but is also required for Aurora B activity...
October 10, 2016: Journal of Cell Biology
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