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Regulation of Cdc42 and Its Effectors in Epithelial Morphogenesis Franck Pichaud1,2,*, Rhian F
© 2019. Published by The Company of Biologists Ltd | Journal of Cell Science (2019) 132, jcs217869. doi:10.1242/jcs.217869 REVIEW SUBJECT COLLECTION: ADHESION Regulation of Cdc42 and its effectors in epithelial morphogenesis Franck Pichaud1,2,*, Rhian F. Walther1 and Francisca Nunes de Almeida1 ABSTRACT An overview of Cdc42 Cdc42 – a member of the small Rho GTPase family – regulates cell Cdc42 was discovered in yeast and belongs to a large family of small – polarity across organisms from yeast to humans. It is an essential (20 30 kDa) GTP-binding proteins (Adams et al., 1990; Johnson regulator of polarized morphogenesis in epithelial cells, through and Pringle, 1990). It is part of the Ras-homologous Rho subfamily coordination of apical membrane morphogenesis, lumen formation and of GTPases, of which there are 20 members in humans, including junction maturation. In parallel, work in yeast and Caenorhabditis elegans the RhoA and Rac GTPases, (Hall, 2012). Rho, Rac and Cdc42 has provided important clues as to how this molecular switch can homologues are found in all eukaryotes, except for plants, which do generate and regulate polarity through localized activation or inhibition, not have a clear homologue for Cdc42. Together, the function of and cytoskeleton regulation. Recent studies have revealed how Rho GTPases influences most, if not all, cellular processes. important and complex these regulations can be during epithelial In the early 1990s, seminal work from Alan Hall and his morphogenesis. This complexity is mirrored by the fact that Cdc42 can collaborators identified Rho, Rac and Cdc42 as main regulators of exert its function through many effector proteins. -
Design of a Bioactive Small Molecule That Targets R(AUUCU) Repeats in Spinocerebellar Ataxia 10
ARTICLE Received 24 Jun 2015 | Accepted 18 Apr 2016 | Published 1 Jun 2016 DOI: 10.1038/ncomms11647 OPEN Design of a bioactive small molecule that targets r(AUUCU) repeats in spinocerebellar ataxia 10 Wang-Yong Yang1, Rui Gao2, Mark Southern3, Partha S. Sarkar2 & Matthew D. Disney1 RNA is an important target for chemical probes of function and lead therapeutics; however, it is difficult to target with small molecules. One approach to tackle this problem is to identify compounds that target RNA structures and utilize them to multivalently target RNA. Here we show that small molecules can be identified to selectively bind RNA base pairs by probing a library of RNA-focused small molecules. A small molecule that selectively binds AU base pairs informed design of a dimeric compound (2AU-2) that targets the pathogenic RNA, expanded r(AUUCU) repeats, that causes spinocerebellar ataxia type 10 (SCA10) in patient- derived cells. Indeed, 2AU-2 (50 nM) ameliorates various aspects of SCA10 pathology including improvement of mitochondrial dysfunction, reduced activation of caspase 3, and reduction of nuclear foci. These studies provide a first-in-class chemical probe to study SCA10 RNA toxicity and potentially define broadly applicable compounds targeting RNA AU base pairs in cells. 1 Departments of Chemistry and Neuroscience, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA. 2 Mitchell Center for Neurodegenerative Disorders, Department of Neurology, Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas 77555, USA. 3 Informatics Core, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA. Correspondence and requests for materials should be addressed to M.D.D. -
Comprehensive Identification and Characterization of Somatic Copy Number Alterations in Triple‑Negative Breast Cancer
INTERNATIONAL JOURNAL OF ONCOLOGY 56: 522-530, 2020 Comprehensive identification and characterization of somatic copy number alterations in triple‑negative breast cancer ZAIBING LI1,2*, XIAO ZHANG3*, CHENXIN HOU4, YUQING ZHOU4, JUNLI CHEN1, HAOYANG CAI5, YIFENG YE3, JINPING LIU3 and NING HUANG1 1Department of Pathophysiology, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041; 2Department of Pathophysiology, School of Basic Medical Science, Southwest Medical University, Luzhou, Sichuan 646000; 3Department of Breast Surgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 611731; 4West China Medical School, Sichuan University, Chengdu, Sichuan 610041; 5Center of Growth, Metabolism and Aging, Key Laboratory of Bio‑Resources and Eco‑Environment, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, P.R. China Received January 30, 2019; Accepted August 30, 2019 DOI: 10.3892/ijo.2019.4950 Abstract. Triple-negative breast cancer (TNBC) accounts hierarchical clustering of tumors resulted in three main for ~15% of all breast cancer diagnoses each year. Patients subgroups that exhibited distinct CNA profiles, which with TNBC tend to have a higher risk for early relapse and may reveal the heterogeneity of molecular mechanisms in a worse prognosis. TNBC is characterized by extensive TNBC subgroups. These results will extend the molecular somatic copy number alterations (CNAs). However, the DNA understanding of TNBC and will facilitate the discovery of CNA profile of TNBC remains to be extensively investigated. therapeutic and diagnostic target candidates. The present study assessed the genomic profile of CNAs in 201 TNBC samples, aiming to identify recurrent CNAs that Introduction may drive the pathogenesis of TNBC. -
The Borg Family of Cdc42 Effector Proteins Cdc42ep1–5
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Institute of Cancer Research Repository Biochemical Society Transactions (2016) 0 1–8 DOI: 10.1042/BST20160219 1 2 The Borg family of Cdc42 effector proteins 3 4 Cdc42EP1–5 5 6 Aaron J. Farrugia and Fernando Calvo 7 8 Tumour Microenvironment Team, Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW2 6JB, U.K. 9 Correspondence: Fernando Calvo ([email protected]) 10 11 12 13 Despite being discovered more than 15 years ago, the Borg (binder of Rho GTPases) 14 – family of Cdc42 effector proteins (Cdc42EP1 5) remains largely uncharacterised and rela- 15 tively little is known about their structure, regulation and role in development and disease. 16 Recent studies are starting to unravel some of the key functional and mechanistic 17 aspects of the Borg proteins, including their role in cytoskeletal remodelling and signal- 18 ling. In addition, the participation of Borg proteins in important cellular processes such as 19 cell shape, directed migration and differentiation is slowly emerging, directly linking Borgs 20 with important physiological and pathological processes such as angiogenesis, neuro- 21 fi transmission and cancer-associated desmoplasia. Here, we review some of these nd- 22 ings and discuss future prospects. 23 24 25 26 27 28 29 Introduction 30 The Rho GTPase family member Cdc42 regulates a diverse range of cellular functions including cyto- 31 kinesis, cytoskeletal remodelling and cell polarity [1,2]. Like other Rho family members, Cdc42 cycles 32 between two tightly regulated conformational states, a GTP-bound active state and a GDP-bound 33 inactive state [3]. -
Supplemental Information
Supplemental information Dissection of the genomic structure of the miR-183/96/182 gene. Previously, we showed that the miR-183/96/182 cluster is an intergenic miRNA cluster, located in a ~60-kb interval between the genes encoding nuclear respiratory factor-1 (Nrf1) and ubiquitin-conjugating enzyme E2H (Ube2h) on mouse chr6qA3.3 (1). To start to uncover the genomic structure of the miR- 183/96/182 gene, we first studied genomic features around miR-183/96/182 in the UCSC genome browser (http://genome.UCSC.edu/), and identified two CpG islands 3.4-6.5 kb 5’ of pre-miR-183, the most 5’ miRNA of the cluster (Fig. 1A; Fig. S1 and Seq. S1). A cDNA clone, AK044220, located at 3.2-4.6 kb 5’ to pre-miR-183, encompasses the second CpG island (Fig. 1A; Fig. S1). We hypothesized that this cDNA clone was derived from 5’ exon(s) of the primary transcript of the miR-183/96/182 gene, as CpG islands are often associated with promoters (2). Supporting this hypothesis, multiple expressed sequences detected by gene-trap clones, including clone D016D06 (3, 4), were co-localized with the cDNA clone AK044220 (Fig. 1A; Fig. S1). Clone D016D06, deposited by the German GeneTrap Consortium (GGTC) (http://tikus.gsf.de) (3, 4), was derived from insertion of a retroviral construct, rFlpROSAβgeo in 129S2 ES cells (Fig. 1A and C). The rFlpROSAβgeo construct carries a promoterless reporter gene, the β−geo cassette - an in-frame fusion of the β-galactosidase and neomycin resistance (Neor) gene (5), with a splicing acceptor (SA) immediately upstream, and a polyA signal downstream of the β−geo cassette (Fig. -
MOLECULAR G EN ETICS of the January, HUMAN X
MOLECULAR GENETICS OF THE HUMAN X CHROMOSOME BY DAVID ANDREW HARTLEY a thesis submitted for the degree of Doctor of Philosophy in the University of London January, 1984 Department of Biochemistry St. Mary's Hospital Medical School London W2 IPG 1 Abstract The comparison of frequency of recombination with chromosomal position has been possible only in Drosophila melanogaster but it is known that chiasma distribution in the human male (and many other species) is not random. A number of cloned DNA sequences have been isolated from a library enriched for X chromosome DNA by flow cytometry. The chromosomal loci complementary to these cloned sequences have been investigated by hybridization to somatic cell hybrid DNAs and by direct hybridization 1 in situ' to human metaphase spreads. In this way a cytological map of the X chromosome defined by DNA sequence markers has been constructed. Genetic distances complementary to the physical separations determined have been estimated by determining the segregation of DNA sequence variants at each locus in three generation p e d i g r e e s . The sequence variation is manifested as altered restriction fragment migration in agarose gels. It is intimated from these analyses that the pattern of recombination frequency along the human X chromosome mirrors the non-random chiasma distribution seen along male autosomes. This supports the chiasmatype theory and has implications on the availability of genetic components of the X chromosome to recombination. 2 To ray father in the hope of convincing him of life outside o f E.coli 3 ...scientists getting their kicks when deadly disease can do as it pleases results ain't hard to predict Gil Scott-Heron 4 Ack n ov; ledgements To my supervisor Bob Williamson for support. -
Reproduction
REPRODUCTIONRESEARCH Focus on Mammalian Embryogenomics Molecular and subcellular characterisation of oocytes screened for their developmental competence based on glucose-6-phosphate dehydrogenase activity Helmut Torner2, Nasser Ghanem, Christina Ambros2, Michael Ho¨lker, Wolfgang Tomek2, Chirawath Phatsara, Hannelore Alm2, Marc-Andre´ Sirard1, Wilhelm Kanitz2, Karl Schellander and Dawit Tesfaye Animal Breeding and Husbandry Group, Department of Animal Breeding and Husbandry, Institute of Animal Science, University of Bonn, Endenicher allee 15, 53115 Bonn, Germany, 1De´partement des Sciences Animales, Centre de Recherche en Biologie de la Reproduction, Universite´ Laval, Pav. Comtois, Laval, Sainte-Foy, Que´bec, G1K 7P4, Canada and 2Research Institute for the Biology of Farm Animals, Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany Correspondence should be addressed to D Tesfaye; Email: [email protected] Abstract Oocyte selection based on glucose-6-phosphate dehydrogenase (G6PDH) activity has been successfully used to differentiate between competent and incompetent bovine oocytes. However, the intrinsic molecular and subcellular characteristics of these oocytes have not yet been investigated. Here, we aim to identify molecular and functional markers associated with oocyte developmental potential when selected based on G6PDH activity. Immature compact cumulus–oocyte complexes were stained with brilliant cresyl blue (BCB) for 90 min. Based on K C their colouration, oocytes were divided into BCB (colourless cytoplasm, high G6PDH activity) and BCB (coloured cytoplasm, low G6PDH activity). The chromatin configuration of the nucleus and the mitochondrial activityof oocytes were determined by fluorescence labelling and photometric measurement. The abundance and phosphorylation pattern of protein kinases Akt and MAP were estimated by Western blot C K analysis. -
Characterizing Humidity, Sex, and B-Cell Gene Regulation in Fungal Allergic
CHARACTERIZING HUMIDITY, SEX, AND B-CELL GENE REGULATION IN FUNGAL ALLERGIC ASTHMA A Thesis Submitted to the Graduate Faculty of the North Dakota State University of Agriculture and Applied Science By Emma Claire Kusick In Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Major Department: Microbiological Sciences April 2020 Fargo, North Dakota North Dakota State University Graduate School Title CHARACTERIZING HUMIDITY, SEX, AND B-CELL GENE REGULATION IN FUNGAL ALLERGIC ASTHMA By EMMA KUSICK The Supervisory Committee certifies that this disquisition complies with North Dakota State University’s regulations and meets the accepted standards for the degree of MASTER OF SCIENCE SUPERVISORY COMMITTEE: Dr. Jane Schuh Chair Dr. Teresa Bergholz Dr. Penelope Gibbs Dr. Sumit Ghosh Approved: 5/4/2020 Dr. John McEvoy Date Department Chair ABSTRACT Asthma is a debilitating lung disease that affects nearly 300 million people worldwide. Environments with high humidity and subsequent mold exposure often trigger allergic asthma. Sex differences have been reported in the incidence, prevalence, and severity of asthma. B-lymphocytes are recruited in high numbers to the allergic lung in response to the inhalation of Aspergillus fumigatus mold spores (conidia). In this work, we used a mouse model of allergic fungal asthma to assess environmental humidity, sex, and B-lymphocytes in an inhalational model of allergic fungal asthma. Our results showed that animals sensitized in low humidity conditions had no airway hyperresponsiveness (AHR), inflammation, but an increase in IgG3 antibody production. Males weighed more than females, female mice had more fibrosis and produced more IgG3 Ab, but sex showed no impact on low humidity. -
CDC42EP4 (NM 012121) Human Tagged ORF Clone Product Data
OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for RC201162L3 CDC42EP4 (NM_012121) Human Tagged ORF Clone Product data: Product Type: Expression Plasmids Product Name: CDC42EP4 (NM_012121) Human Tagged ORF Clone Tag: Myc-DDK Symbol: CDC42EP4 Synonyms: BORG4; CEP4; KAIA1777 Vector: pLenti-C-Myc-DDK-P2A-Puro (PS100092) E. coli Selection: Chloramphenicol (34 ug/mL) Cell Selection: Puromycin ORF Nucleotide The ORF insert of this clone is exactly the same as(RC201162). Sequence: Restriction Sites: SgfI-MluI Cloning Scheme: ACCN: NM_012121 ORF Size: 1068 bp This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 CDC42EP4 (NM_012121) Human Tagged ORF Clone – RC201162L3 OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene. RefSeq: NM_012121.4 RefSeq Size: 3121 bp RefSeq ORF: 1071 bp Locus ID: 23580 UniProt ID: Q9H3Q1, B2R6D8 Domains: PBD MW: 38 kDa Gene Summary: The product of this gene is a member of the CDC42-binding protein family. -
Molecular Analyses of Malignant Pleural Mesothelioma
Molecular Analyses of Malignant Pleural Mesothelioma Shir Kiong Lo National Heart and Lung Institute Imperial College Dovehouse Street London SW3 6LY A thesis submitted for MD (Res) Faculty of Medicine, Imperial College London 2016 1 Abstract Malignant pleural mesothelioma (MPM) is an aggressive cancer that is strongly associated with asbestos exposure. Majority of patients with MPM present with advanced disease and the treatment paradigm mainly involves palliative chemotherapy and best supportive care. The current chemotherapy options are limited and ineffective hence there is an urgent need to improve patient outcomes. This requires better understanding of the genetic alterations driving MPM to improve diagnostic, prognostic and therapeutic strategies. This research aims to gain further insights in the pathogenesis of MPM by exploring the tumour transcriptional and mutational profiles. We compared gene expression profiles of 25 MPM tumours and 5 non-malignant pleura. This revealed differentially expressed genes involved in cell migration, invasion, cell cycle and the immune system that contribute to the malignant phenotype of MPM. We then constructed MPM-associated co-expression networks using weighted gene correlation network analysis to identify clusters of highly correlated genes. These identified three distinct molecular subtypes of MPM associated with genes involved in WNT and TGF-ß signalling pathways. Our results also revealed genes involved in cell cycle control especially the mitotic phase correlated significantly with poor prognosis. Through exome analysis of seven paired tumour/blood and 29 tumour samples, we identified frequent mutations in BAP1 and NF2. Additionally, the mutational profile of MPM is enriched with genes encoding FAK, MAPK and WNT signalling pathways. -
Autosomal Dominant Cerebellar Ataxia Type I: a Review of the Phenotypic and Genotypic Characteristics
Whaley et al. Orphanet Journal of Rare Diseases 2011, 6:33 http://www.ojrd.com/content/6/1/33 REVIEW Open Access Autosomal dominant cerebellar ataxia type I: A review of the phenotypic and genotypic characteristics Nathaniel Robb Whaley1,2, Shinsuke Fujioka2 and Zbigniew K Wszolek2* Abstract Type I autosomal dominant cerebellar ataxia (ADCA) is a type of spinocerebellar ataxia (SCA) characterized by ataxia with other neurological signs, including oculomotor disturbances, cognitive deficits, pyramidal and extrapyramidal dysfunction, bulbar, spinal and peripheral nervous system involvement. The global prevalence of this disease is not known. The most common type I ADCA is SCA3 followed by SCA2, SCA1, and SCA8, in descending order. Founder effects no doubt contribute to the variable prevalence between populations. Onset is usually in adulthood but cases of presentation in childhood have been reported. Clinical features vary depending on the SCA subtype but by definition include ataxia associated with other neurological manifestations. The clinical spectrum ranges from pure cerebellar signs to constellations including spinal cord and peripheral nerve disease, cognitive impairment, cerebellar or supranuclear ophthalmologic signs, psychiatric problems, and seizures. Cerebellar ataxia can affect virtually any body part causing movement abnormalities. Gait, truncal, and limb ataxia are often the most obvious cerebellar findings though nystagmus, saccadic abnormalities, and dysarthria are usually associated. To date, 21 subtypes have been identified: SCA1-SCA4, SCA8, SCA10, SCA12-SCA14, SCA15/16, SCA17-SCA23, SCA25, SCA27, SCA28 and dentatorubral pallidoluysian atrophy (DRPLA). Type I ADCA can be further divided based on the proposed pathogenetic mechanism into 3 subclasses: subclass 1 includes type I ADCA caused by CAG repeat expansions such as SCA1-SCA3, SCA17, and DRPLA, subclass 2 includes trinucleotide repeat expansions that fall outside of the protein-coding regions of the disease gene including SCA8, SCA10 and SCA12. -
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bioRxiv preprint doi: https://doi.org/10.1101/751933; this version posted September 6, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Genetics and Pathway Analysis of Normative Cognitive Variation 2 in the Philadelphia Neurodevelopmental Cohort 3 4 Authors: 5 Shraddha Pai1, Shirley Hui1, Philipp Weber2, Owen Whitley1,3, Peipei Li4,5, Viviane Labrie4,5, Jan 6 Baumbach2,6, Gary D Bader1,3,7,8 7 8 Affiliations: 9 1. The Donnelly Centre, University of Toronto, Toronto, Canada 10 2. Department of Mathematics and Computer Science, University of Southern Denmark, Odense, 11 Denmark 12 3. Department of Molecular Genetics, University of Toronto, Toronto, Canada 13 4. Center for Neurodegenerative Science, Van Andel Research Institute, Grand Rapids, MI, USA 14 5. Division of Psychiatry and Behavioral Medicine, College of Human Medicine, Michigan State 15 University, Grand Rapids, MI, USA 16 6. TUM School of Life Sciences Weihenstephan, Technical University of Munich, Munich, Germany. 17 7. Department of Computer Science, University of Toronto, Toronto, Canada 18 8. The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada 19 20 21 22 * [email protected] 23 24 Page 1 of 23 bioRxiv preprint doi: https://doi.org/10.1101/751933; this version posted September 6, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.