DOCSLIB.ORG

Finding Pathogenic Nssnp's and Their Structural Effect on COPS2 Using Molecular Dynamic Approach

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Finding Pathogenic Nssnp's and Their Structural Effect on COPS2 Using Molecular Dynamic Approach bioRxiv preprint doi: https://doi.org/10.1101/2020.10.12.333252; this version posted October 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Finding Pathogenic nsSNP's and their structural effect on COPS2 using Molecular Dynamic Approach Ashish Malik1, Kajal Pande1, Abhishek Kumar1, Alekhya Vemula1, Madhuri R1 Vivek Chandramohan2* 1) BiSEP, Bioinformatics and Big data analytics, Department of Biotechnology, Siddaganga Institute of Technology, Tumkur. 2) Department of Biotechnology, Siddaganga Institute of Technology, Tumkur. Corresponding author: Mr. Vivek Chandramohan (M.Sc) Assistant Professor, Backup Coordinator for Biotechnology Skill Enhancement Programme (BiSEP) Department of Biotechnology, Siddaganga Institute of Technology, Tumkur - 572103 bioRxiv preprint doi: https://doi.org/10.1101/2020.10.12.333252; this version posted October 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract COP9 Signalosome Subunit 2 is a highly conserved multiprotein complex which is involved in the cellular process and developmental process. It is one of the essential components in the COP9 Signalosome Complex (CSN). It is also involved in neuronal differentiation interacting with NIF3L1. The gene involved in neuronal differentiation is negatively regulated due to the transcription co-repressor interaction of NIF3L1 with COPS2. In the present study, we have evaluated the outcome for 90 non- synonymous single nucleotide polymorphisms (nsSNP’s) in COPS2 gene through computational tools. After the analysis, 4 SNP’s (S120C, N144S, Y159H, R173C) were found to be deleterious. The native and mutated structures were prepared using discovery studio and docked to check the interactions with NIF3L1.On the basis of ZDOCK score the top 3 mutations (N144S, Y159H, R173C) were screened out. Further to analyze the effect of amino acid substitution on the molecular structure of protein Molecular Dynamics simulation was carried out. Analysis based on RMSD, RMSF, RG, H-bond showed a significant deviation in the graph, which demonstrated conformation change and instability compared to the wild structure. As it is known mutations in COPS2 gene can disrupt the normal activity of the CSN2 protein which may cause neuronal differentiation. Our results showed N144S, Y159H and R173C mutations are to be more pathogenic and may cause disease KEYWORDS: Homology modeling, MD simulation, protein-protein docking, single nucleotide polymorphism bioRxiv preprint doi: https://doi.org/10.1101/2020.10.12.333252; this version posted October 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Introduction COP9 Signalosome Subunit 2 (COPS2) is one of the important component of COP9 signalosome complex (CSN) that exists in eukaryotes. There are many synonyms that are in use for COPS2 like, TRIP15, SGN2 and CSN2. CSN is a conserved multi-protein complex (CSN1 to CSN8) that has seemingly evolved in parallel with the ubiquitin- proteasome system [1]. In a eukaryotic organism, CSN complex is associated to be as an essential protein complex that is involved in multiple cellular and developmental processes. Any deletion of CSN subunits can cause serious changes in the gene articulation profile and can cause early lethality of the multicellular organism [2–4]. The CSN complex was initially discovered as a eliminator of photomorphogenesis in Arabidopsis thaliana [1]. The eight mammalian complex subunits are classified according to the molecular weight S1 (57 kDa) to S8 (22 kDa) [5]. COP9 signalosome complex was considered as contaminating component in 26S proteasome preparation from erythrocytes by Seegeret al. and they later separated the two complexes [6]. The complex was cleansed biochemically from pig spleen by [5]. The CSN complex is a crucial regulator of ubiquitin (Ubl) conjugation pathway by mediating the deneddylation of the cullin subunits [7]. The subunits of CSN shows notable sequence homologies to elements of the lid subcomplex of the 26S proteasome which is formed by the assembly of 19S proteasome regulatory complex with the 20S catalytic core [8] and the translation initiation complex eIF3[6]. In eukaryotes, the eIF3 complex is involved in protein synthesis and ribosome biogenesis and it serves as a binding site for assembling other eIFsduring translation initiation [9]. Initially, Trip15/CSN2 was recognized as a thyroid hormone receptor (TR)-interacting protein and acts as a transcriptional corepressor [5]. In yeast two-hybrid screen, Trip15 is reported to interact with the thyroid hormone receptor and the retinoic acid X receptor in a ligand- dependent manner and the N-terminal region of subunit 2 (S2) is identical to Trip15[10]. Even though the mutual interaction between Trip15-nuclear receptor complex and CSN complex is obscure, indicating that the CSN complex assists in several signal transduction pathways [11]. COPS2 is a 51,597 Da protein with a sequence length of 443 amino acids. It also dissipates the interaction between OCT4 and CDK1 by segregating OCT4 and forming a COPS2/CDK1 complex thus hindering the inhibitory effect of OCT4 on CDK1 activation[12]. COPS2 (CSN2) shows transcriptional corepressor activity. The protein-protein interaction occurring inside the CSN complex (21-24) is stabilized by the proteasome COP9 complex initiation factor domain 3 in the C-terminal region of the Trip15/CSN whereas the N-terminal region of CSN2 has been reported to be adequate for the effector function of Trip15/CSN2. Trip15/CSN2 is associated with its binding partner through N terminal (Akiyama et al. 2003). The CS2 domain region (1-275) mediates interaction with NIF3L1, which is a highly conserved gene from bacteria to human. Although it is known that the mouse NIF3L1 gene encodes for a cytoplasmic protein (376 amino acids) and the expression of the protein was detected through mouse embryonic development, still the real-biological function of Nif3l1 is still unknown. In order to study the association of Trip15/CSN2 with Nif3l1, a wet lab study was carried out by Akiyama and co to exhibit that Nif3l1 protein could be translocated into nuclei by Trip15/CSN2 and it is involved in neural differentiation through down-regulation of Oct-¾ transcript which suppresses neurogenic genes. By the result of the study carried out by Akiyama and co, it seems likely that Nif3l1 and Trip15/CSN2 are associated and play an important role in neural differentiation [11]. NIF3L1 is engaged in retinoic acid primed neural differentiation of P19 embryonic carcinoma cells through cooperation with CSN2 [11]. In the complex COPS2 bioRxiv preprint doi: https://doi.org/10.1101/2020.10.12.333252; this version posted October 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. interacts with COPS1, COPS4, COPS5, COPS6, COPS7A or COPS7B directly[8, 13]. Notably, it also interacts with the ligand binding region of thyroid receptor, CUL1 and CUL2 [8]. Single nucleotide polymorphism is known as the most common type of variants occurring in human genome due to single nucleotide substitution [14, 15]. Non- synonymous SNPs can cause 50% of the disease that is related to inheritance [16]. SNPs can also disrupt the normal function of the protein and denature the structure by changing the stability, folding pattern and ligand binding site [16]. The amino acid changes at key sites within a protein may result in series of conformation changes, including the disintegration of salt bridges, alteration of interaction network, disruption in hydrogen bonds, which in turn may disturb the energy landscape [17]. Protein structure modeling methods have been extensively used for predicting the effects of disease-causing mutation on protein stability and protein-protein interaction [18]. The aim of the present study was to identify the most deleterious nsSNPs within COPS2 gene and their effect on protein structure. Protein-protein docking was done in order to determine the effect of amino acid substitution on the structure of COPS2 which will affect the interaction with Nif3l1. To check the interaction of COPS2 with NIF3L1, protein-protein docking was done. The mutations which are identified to have a positive interrelationship with pathogenicity and have high ZDOCK score were further investigated using MD simulations to determine the changes on its time-dependent physiological affinities and biochemical pathway alterations (Sinha et al. 2018). We utilized RMSD, RMSF, H-Bond, Gyration, in order to examine the motion trajectory and atomic interactions of native and mutant protein structures. Alteration in the protein structure can lead to a change in the binding site of protein which may affect the interacting proteins of COPS2 that may cause disease. bioRxiv preprint doi: https://doi.org/10.1101/2020.10.12.333252; this version posted October 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Materials and Methods SNPs Data Collection The data of human COPS2 gene were obtained from the NCBI database of dbSNP (www.ncbi.nlm.nih.gov/snp),
Load more

Recommended publications
  • Old Data and Friends Improve with Age: Advancements with the Updated Tools of Genenetwork
    bioRxiv preprint doi: https://doi.org/10.1101/2021.05.24.445383; this version posted May 25, 2021. 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. Old data and friends improve with age: Advancements with the updated tools of GeneNetwork Alisha Chunduri1, David G. Ashbrook2 1Department of Biotechnology, Chaitanya Bharathi Institute of Technology, Hyderabad 500075, India 2Department of Genetics, Genomics and Informatics, University of Tennessee Health Science Center, Memphis, TN 38163, USA Abstract Understanding gene-by-environment interactions is important across biology, particularly behaviour. Families of isogenic strains are excellently placed, as the same genome can be tested in multiple environments. The BXD’s recent expansion to 140 strains makes them the largest family of murine isogenic genomes, and therefore give great power to detect QTL. Indefinite reproducible genometypes can be leveraged; old data can be reanalysed with emerging tools to produce novel biological insights. To highlight the importance of reanalyses, we obtained drug- and behavioural-phenotypes from Philip et al. 2010, and reanalysed their data with new genotypes from sequencing, and new models (GEMMA and R/qtl2). We discover QTL on chromosomes 3, 5, 9, 11, and 14, not found in the original study. We narrowed down the candidate genes based on their ability to alter gene expression and/or protein function, using cis-eQTL analysis, and variants predicted to be deleterious. Co-expression analysis (‘gene friends’) and human PheWAS were used to further narrow candidates.
    [Show full text]
  • Analysis of Trans Esnps Infers Regulatory Network Architecture
    Analysis of trans eSNPs infers regulatory network architecture Anat Kreimer Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2014 © 2014 Anat Kreimer All rights reserved ABSTRACT Analysis of trans eSNPs infers regulatory network architecture Anat Kreimer eSNPs are genetic variants associated with transcript expression levels. The characteristics of such variants highlight their importance and present a unique opportunity for studying gene regulation. eSNPs affect most genes and their cell type specificity can shed light on different processes that are activated in each cell. They can identify functional variants by connecting SNPs that are implicated in disease to a molecular mechanism. Examining eSNPs that are associated with distal genes can provide insights regarding the inference of regulatory networks but also presents challenges due to the high statistical burden of multiple testing. Such association studies allow: simultaneous investigation of many gene expression phenotypes without assuming any prior knowledge and identification of unknown regulators of gene expression while uncovering directionality. This thesis will focus on such distal eSNPs to map regulatory interactions between different loci and expose the architecture of the regulatory network defined by such interactions. We develop novel computational approaches and apply them to genetics-genomics data in human. We go beyond pairwise interactions to define network motifs, including regulatory modules and bi-fan structures, showing them to be prevalent in real data and exposing distinct attributes of such arrangements. We project eSNP associations onto a protein-protein interaction network to expose topological properties of eSNPs and their targets and highlight different modes of distal regulation.
    [Show full text]
  • ACE2 Interaction Networks in COVID-19: a Physiological Framework for Prediction of Outcome in Patients with Cardiovascular Risk Factors
    Journal of Clinical Medicine Article ACE2 Interaction Networks in COVID-19: A Physiological Framework for Prediction of Outcome in Patients with Cardiovascular Risk Factors Zofia Wicik 1,2 , Ceren Eyileten 2, Daniel Jakubik 2,Sérgio N. Simões 3, David C. Martins Jr. 1, Rodrigo Pavão 1, Jolanta M. Siller-Matula 2,4,* and Marek Postula 2 1 Centro de Matemática, Computação e Cognição, Universidade Federal do ABC, Santo Andre 09606-045, Brazil; zofi[email protected] (Z.W.); [email protected] (D.C.M.J.); [email protected] (R.P.) 2 Department of Experimental and Clinical Pharmacology, Medical University of Warsaw, Center for Preclinical Research and Technology CEPT, 02-091 Warsaw, Poland; [email protected] (C.E.); [email protected] (D.J.); [email protected] (M.P.) 3 Federal Institute of Education, Science and Technology of Espírito Santo, Serra, Espírito Santo 29056-264, Brazil; [email protected] 4 Department of Internal Medicine II, Division of Cardiology, Medical University of Vienna, 1090 Vienna, Austria * Correspondence: [email protected]; Tel.: +43-1-40400-46140; Fax: +43-1-40400-42160 Received: 9 October 2020; Accepted: 17 November 2020; Published: 21 November 2020 Abstract: Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (coronavirus disease 2019; COVID-19) is associated with adverse outcomes in patients with cardiovascular disease (CVD). The aim of the study was to characterize the interaction between SARS-CoV-2 and Angiotensin-Converting Enzyme 2 (ACE2) functional networks with a focus on CVD. Methods: Using the network medicine approach and publicly available datasets, we investigated ACE2 tissue expression and described ACE2 interaction networks that could be affected by SARS-CoV-2 infection in the heart, lungs and nervous system.
    [Show full text]
  • Transcriptome Analyses of Rhesus Monkey Pre-Implantation Embryos Reveal A
    Downloaded from genome.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press Transcriptome analyses of rhesus monkey pre-implantation embryos reveal a reduced capacity for DNA double strand break (DSB) repair in primate oocytes and early embryos Xinyi Wang 1,3,4,5*, Denghui Liu 2,4*, Dajian He 1,3,4,5, Shengbao Suo 2,4, Xian Xia 2,4, Xiechao He1,3,6, Jing-Dong J. Han2#, Ping Zheng1,3,6# Running title: reduced DNA DSB repair in monkey early embryos Affiliations: 1 State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China 2 Key Laboratory of Computational Biology, CAS Center for Excellence in Molecular Cell Science, Collaborative Innovation Center for Genetics and Developmental Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China 3 Yunnan Key Laboratory of Animal Reproduction, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China 4 University of Chinese Academy of Sciences, Beijing, China 5 Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, Yunnan 650204, China 6 Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, China * Xinyi Wang and Denghui Liu contributed equally to this work 1 Downloaded from genome.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press # Correspondence: Jing-Dong J. Han, Email: [email protected]; Ping Zheng, Email: [email protected] Key words: rhesus monkey, pre-implantation embryo, DNA damage 2 Downloaded from genome.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press ABSTRACT Pre-implantation embryogenesis encompasses several critical events including genome reprogramming, zygotic genome activation (ZGA) and cell fate commitment.
    [Show full text]
  • Seq2pathway Vignette
    seq2pathway Vignette Bin Wang, Xinan Holly Yang, Arjun Kinstlick May 19, 2021 Contents 1 Abstract 1 2 Package Installation 2 3 runseq2pathway 2 4 Two main functions 3 4.1 seq2gene . .3 4.1.1 seq2gene flowchart . .3 4.1.2 runseq2gene inputs/parameters . .5 4.1.3 runseq2gene outputs . .8 4.2 gene2pathway . 10 4.2.1 gene2pathway flowchart . 11 4.2.2 gene2pathway test inputs/parameters . 11 4.2.3 gene2pathway test outputs . 12 5 Examples 13 5.1 ChIP-seq data analysis . 13 5.1.1 Map ChIP-seq enriched peaks to genes using runseq2gene .................... 13 5.1.2 Discover enriched GO terms using gene2pathway_test with gene scores . 15 5.1.3 Discover enriched GO terms using Fisher's Exact test without gene scores . 17 5.1.4 Add description for genes . 20 5.2 RNA-seq data analysis . 20 6 R environment session 23 1 Abstract Seq2pathway is a novel computational tool to analyze functional gene-sets (including signaling pathways) using variable next-generation sequencing data[1]. Integral to this tool are the \seq2gene" and \gene2pathway" components in series that infer a quantitative pathway-level profile for each sample. The seq2gene function assigns phenotype-associated significance of genomic regions to gene-level scores, where the significance could be p-values of SNPs or point mutations, protein-binding affinity, or transcriptional expression level. The seq2gene function has the feasibility to assign non-exon regions to a range of neighboring genes besides the nearest one, thus facilitating the study of functional non-coding elements[2]. Then the gene2pathway summarizes gene-level measurements to pathway-level scores, comparing the quantity of significance for gene members within a pathway with those outside a pathway.
    [Show full text]
  • Epigenetic Mechanisms of Lncrnas Binding to Protein in Carcinogenesis
    cancers Review Epigenetic Mechanisms of LncRNAs Binding to Protein in Carcinogenesis Tae-Jin Shin, Kang-Hoon Lee and Je-Yoel Cho * Department of Biochemistry, BK21 Plus and Research Institute for Veterinary Science, School of Veterinary Medicine, Seoul National University, Seoul 08826, Korea; [email protected] (T.-J.S.); [email protected] (K.-H.L.) * Correspondence: [email protected]; Tel.: +82-02-800-1268 Received: 21 September 2020; Accepted: 9 October 2020; Published: 11 October 2020 Simple Summary: The functional analysis of lncRNA, which has recently been investigated in various fields of biological research, is critical to understanding the delicate control of cells and the occurrence of diseases. The interaction between proteins and lncRNA, which has been found to be a major mechanism, has been reported to play an important role in cancer development and progress. This review thus organized the lncRNAs and related proteins involved in the cancer process, from carcinogenesis to metastasis and resistance to chemotherapy, to better understand cancer and to further develop new treatments for it. This will provide a new perspective on clinical cancer diagnosis, prognosis, and treatment. Abstract: Epigenetic dysregulation is an important feature for cancer initiation and progression. Long non-coding RNAs (lncRNAs) are transcripts that stably present as RNA forms with no translated protein and have lengths larger than 200 nucleotides. LncRNA can epigenetically regulate either oncogenes or tumor suppressor genes. Nowadays, the combined research of lncRNA plus protein analysis is gaining more attention. LncRNA controls gene expression directly by binding to transcription factors of target genes and indirectly by complexing with other proteins to bind to target proteins and cause protein degradation, reduced protein stability, or interference with the binding of other proteins.
    [Show full text]
  • A Compendium of Co-Regulated Protein Complexes in Breast Cancer Reveals Collateral Loss Events
    bioRxiv preprint doi: https://doi.org/10.1101/155333; this version posted June 26, 2017. 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. A compendium of co-regulated protein complexes in breast cancer reveals collateral loss events Colm J. Ryan*1, Susan Kennedy1, Ilirjana Bajrami2, David Matallanas1, Christopher J. Lord2 1Systems Biology Ireland, School of Medicine, University College Dublin, Dublin 4, Ireland 2The Breast Cancer Now Toby Robins Breast Cancer Research Centre and CRUK Gene Function Laboratory, The Institute of Cancer Research, London, SW3 6JB, United Kingdom. * Correspondence: [email protected] Summary Protein complexes are responsible for the bulk of activities within the cell, but how their behavior and composition varies across tumors remains poorly understood. By combining proteomic profiles of breast tumors with a large-scale protein-protein interaction network, we have identified a set of 258 high-confidence protein complexes whose subunits have highly correlated protein abundance across tumor samples. We used this set to identify complexes that are reproducibly under- or over- expressed in specific breast cancer subtypes. We found that mutation or deletion of one subunit of a complex was often associated with a collateral reduction in protein expression of additional complex members. This collateral loss phenomenon was evident from proteomic, but not transcriptomic, profiles suggesting post- transcriptional control. Mutation of the tumor suppressor E-cadherin (CDH1) was associated with a collateral loss of members of the adherens junction complex, an effect we validated using an engineered model of E-cadherin loss.
    [Show full text]
  • Probabilistic Models for Collecting, Analyzing, and Modeling Expression Data
    Probabilistic Models for Collecting, Analyzing, and Modeling Expression Data Hai-Son Phuoc Le May 2013 CMU-ML-13-101 Probabilistic Models for Collecting, Analyzing, and Modeling Expression Data Hai-Son Phuoc Le May 2013 CMU-ML-13-101 Machine Learning Department School of Computer Science Carnegie Mellon University Thesis Committee Ziv Bar-Joseph, Chair Christopher Langmead Roni Rosenfeld Quaid Morris Submitted in partial fulfillment of the requirements for the Degree of Doctor of Philosophy. Copyright @ 2013 Hai-Son Le This research was sponsored by the National Institutes of Health under grant numbers 5U01HL108642 and 1R01GM085022, the National Science Foundation under grant num- bers DBI0448453 and DBI0965316, and the Pittsburgh Life Sciences Greenhouse. The views and conclusions contained in this document are those of the author and should not be interpreted as representing the official policies, either expressed or implied, of any sponsoring institution, the U.S. government or any other entity. Keywords: genomics, gene expression, gene regulation, microarray, RNA-Seq, transcriptomics, error correction, comparative genomics, regulatory networks, cross-species, expression database, Gene Expression Omnibus, GEO, orthologs, microRNA, target prediction, Dirichlet Process, Indian Buffet Process, hidden Markov model, immune response, cancer. To Mom and Dad. i Abstract Advances in genomics allow researchers to measure the complete set of transcripts in cells. These transcripts include messenger RNAs (which encode for proteins) and microRNAs, short RNAs that play an important regulatory role in cellular networks. While this data is a great resource for reconstructing the activity of networks in cells, it also presents several computational challenges. These challenges include the data collection stage which often results in incomplete and noisy measurement, developing methods to integrate several experiments within and across species, and designing methods that can use this data to map the interactions and networks that are activated in specific conditions.
    [Show full text]
  • Cops5 Safeguards Genomic Stability of Embryonic Stem Cells Through Regulating Cellular Metabolism and DNA Repair
    Cops5 safeguards genomic stability of embryonic stem cells through regulating cellular metabolism and DNA repair Peng Lia, Lulu Gaoa, Tongxi Cuia, Weiyu Zhanga, Zixin Zhaoa, and Lingyi Chena,1 aState Key Laboratory of Medicinal Chemical Biology, Collaborative Innovation Center of Tianjin for Medical Epigenetics, Collaborative Innovation Center for Biotherapy, Tianjin Key Laboratory of Protein Sciences, National Demonstration Center for Experimental Biology Education and College of Life Sciences, Nankai University, 300071 Tianjin, China Edited by Janet Rossant, Hospital for Sick Children, University of Toronto, Toronto, Canada, and approved December 24, 2019 (received for review August 29, 2019) The highly conserved COP9 signalosome (CSN), composed of 8 transiently expressed in about 5% of ESCs at a given time, subunits (Cops1 to Cops8), has been implicated in pluripotency promotes rapid telomere elongation by telomere recombination maintenance of human embryonic stem cells (ESCs). Yet, the mech- and regulates genomic stability (11). Induced by genotoxic stress, anism for the CSN to regulate pluripotency remains elusive. We Filia stimulates the PARP1 activity and relocates from centro- previously showed that Cops2, independent of the CSN, is essential somes to DNA damage sites and mitochondria to regulate DDR for the pluripotency maintenance of mouse ESCs. In this study, we and apoptosis (12). Sall4, a pluripotency transcription factor, set out to investigate how Cops5 and Cops8 regulate ESC differ- facilitates the ataxia telangiectasia-mutated activation in re- entiation and tried to establish Cops5 and Cops8 knockout (KO) sponse to DSBs (13). To minimize the ROS-induced genomic ESC lines by CRISPR/Cas9. To our surprise, no Cops5 KO ESC clones DNA damage, ESCs produce lower levels of mitochondrial ROS were identified out of 127 clones, while three Cops8 KO ESC lines and express higher levels of antioxidants than differentiated cells were established out of 70 clones.
    [Show full text]
  • Viewed Graphically with a Java Graphical User Interface (GUI), Allowing Users to Evaluate Contig Sequence Quality and Predict Snps
    BMC Research Notes BioMed Central Short Report Open Access BEAP: The BLAST Extension and Alignment Program- a tool for contig construction and analysis of preliminary genome sequence James E Koltes†, Zhi-Liang Hu†, Eric Fritz and James M Reecy* Address: Department of Animal Science, Iowa State University, Ames, Iowa 50011-3150, USA Email: James E Koltes - [email protected]; Zhi-Liang Hu - [email protected]; Eric Fritz - [email protected]; James M Reecy* - [email protected] * Corresponding author †Equal contributors Published: 22 January 2009 Received: 29 October 2008 Accepted: 22 January 2009 BMC Research Notes 2009, 2:11 doi:10.1186/1756-0500-2-11 This article is available from: http://www.biomedcentral.com/1756-0500/2/11 © 2009 Reecy et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Fine-mapping projects require a high density of SNP markers and positional candidate gene sequences. In species with incomplete genomic sequence, the DNA sequences needed to generate markers for fine-mapping within a linkage analysis confidence interval may be available but may not have been assembled. To manually piece these sequences together is laborious and costly. Moreover, annotation and assembly of short, incomplete DNA sequences is time consuming and not always straightforward. Findings: We have created a tool called BEAP that combines BLAST and CAP3 to retrieve sequences and construct contigs for localized genomic regions in species with unfinished sequence drafts.
    [Show full text]
  • Sex Determination: a 'Window' of DAX1 Activity
    Review TRENDS in Endocrinology and Metabolism Vol.15 No.3 April 2004 Sex determination: a ‘window’ of DAX1 activity Louisa M. Ludbrook and Vincent R. Harley Prince Henry’s Institute of Medical Research, PO Box 5152, Clayton, VIC 3168, Australia Traditionally, DAX1 was considered an ‘anti-testis’ gene that are probably important for male sex determination because DAX1 duplications in XY individuals cause have yet to be identified, because some 75% of sex reversal male-to-female sex reversal: dosage-sensitive sex rever- cases remain unexplained genetically [15]. Some progress sal (DSS). In DSS, two active DAX1 genes on one has been made in deciphering the roles and complex X chromosome can abrogate testis formation. By con- relationships of the known sex-determining genes during trast, mutations and deletions of DAX1 cause adrenal gonadogenesis. Here, we describe the emerging role of hypoplasia congenita (AHC). Although AHC patients DAX1 in male testis formation and discuss the possible develop testes, gonadal defects include disorganized molecular mechanisms through which DAX1 regulates testis cords and hypogonadotropic hypogonadism, this pathway. which is not completely restored with gonadotropin or androgen therapy. Recent evidence of XY sex reversal Expression of DAX1 in Dax1-deficient mice strongly supports a role for Dax1 DAX1 RNA expression is restricted to certain tissue types as a ‘pro-testis’ gene. Therefore, perhaps DAX1/Dax1 and is largely coexpressed with SF1, also crucial for both acts within a ‘window’ of activity, outside of which tes- adrenal and gonadal development [16–18]. Based on in tis formation does not occur. Here, we discuss the func- situ hybridization analyses, Sf1 and Dax1 are expressed in tion and possible mechanisms of DAX1 action in male both developing and adult adrenal, gonadal, hypothalamic gonadogenesis.
    [Show full text]
  • Gene Co-Expression Network Mining Using Graph Sparsification
    Gene Co-expression Network Mining Using Graph Sparsification THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Jinchao Di Graduate Program in Electrical and Computer Engineering The Ohio State University 2013 Master’s Examination Committee: Dr. Kun Huang, Advisor Dr. Raghu Machiraju Dr. Yuejie Chi ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! Copyright by Jinchao Di 2013 ! ! Abstract Identifying and analyzing gene modules is important since it can help understand gene function globally and reveal the underlying molecular mechanism. In this thesis, we propose a method combining local graph sparsification and a gene similarity measurement method TOM. This method can detect gene modules from a weighted gene co-expression network more e↵ectively since we set a local threshold which can detect clusters with di↵erent densities. In our algorithm, we only retain one edge for each gene, which can generate more balanced gene clusters. To estimate the e↵ectiveness of our algorithm we use DAVID to functionally evaluate the gene list for each module we detect and compare our results with some well known algorithms. The result of our algorithm shows better biological relevance than the compared methods and the number of the meaningful biological clusters is much larger than other methods, implying we discover some previously missed gene clusters. Moreover, we carry out a robustness test by adding Gaussian noise with di↵erent variance to the expression data. We find that our algorithm is robust to noise. We also find that some hub genes considered as important genes could be artifacts.
    [Show full text]