Technological Breakthroughs Transform Biological Inquiry in Neuroscience
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The Role of Histone H2av Variant Replacement and Histone H4 Acetylation in the Establishment of Drosophila Heterochromatin
The role of histone H2Av variant replacement and histone H4 acetylation in the establishment of Drosophila heterochromatin Jyothishmathi Swaminathan, Ellen M. Baxter, and Victor G. Corces1 Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218, USA Activation and repression of transcription in eukaryotes involve changes in the chromatin fiber that can be accomplished by covalent modification of the histone tails or the replacement of the canonical histones with other variants. Here we show that the histone H2A variant of Drosophila melanogaster, H2Av, localizes to the centromeric heterochromatin, and it is recruited to an ectopic heterochromatin site formed by a transgene array. His2Av behaves genetically as a PcG gene and mutations in His2Av suppress position effect variegation (PEV), suggesting that this histone variant is required for euchromatic silencing and heterochromatin formation. His2Av mutants show reduced acetylation of histone H4 at Lys 12, decreased methylation of histone H3 at Lys 9, and a reduction in HP1 recruitment to the centromeric region. H2Av accumulation or histone H4 Lys 12 acetylation is not affected by mutations in Su(var)3-9 or Su(var)2-5. The results suggest an ordered cascade of events leading to the establishment of heterochromatin and requiring the recruitment of the histone H2Av variant followed by H4 Lys 12 acetylation as necessary steps before H3 Lys 9 methylation and HP1 recruitment can take place. [Keywords: Chromatin; silencing; transcription; histone; nucleus] Received September 8, 2004; revised version accepted November 4, 2004. The basic unit of chromatin is the nucleosome, which is guchi et al. 2004). The role of histone variants, and spe- made up of 146 bp of DNA wrapped around a histone cially those of H3 and H2A, in various nuclear processes octamer composed of two molecules each of the histones has been long appreciated (Wolffe and Pruss 1996; Ah- H2A, H2B, H3, and H4. -
James Spudich, Ph.D. the Myosin Mesa and Beyond: on the Underlying Molecular Basis of Hyper-Contractility Caused by Hypertrophic Card
JAMES SPUDICH, PH.D. THE MYOSIN MESA AND BEYOND: ON THE UNDERLYING MOLECULAR BASIS OF HYPER-CONTRACTILITY CAUSED BY HYPERTROPHIC CARD APRIL 6, 2017 3:00 P.M. 208 LIGHT HALL SPONSORED BY: THE DEPARTMENT OF CELL AND DEVELOPMENTAL BIOLOGY Upcoming Discovery Lecture: ERIC GOUAUX, PH.D. Senior Scientist, Vollum Institute Jennifer and Bernard Lacroute Term Chair in Neuroscience Research Investigator, HHMI April 20, 2017 208 Light Hall / 4:00 P.M. 1190-4347-Institution-Discovery Lecture Series-Spudich-BK-CH.indd 1 3/21/17 11:11 AM THE MYOSIN MESA AND BEYOND: ON THE UNDERLYING MOLECULAR BASIS OF HYPER-CONTRACTILITY CAUSED BY JAMES SPUDICH, PH.D. STANFORD UNIVERSITY SCHOOL OF MEDICINE DEPARTMENT OF BIOCHEMISTRY THE MYOSIN MESA AND BEYOND: DOUGLASS M. AND NOLA LEISHMAN PROFESSOR OF ON THE UNDERLYING MOLECULAR BASIS CARDIOVASCULAR DISEASE OF HYPER-CONTRACTILITY CAUSED BY MEMBER, NATIONAL ACADEMY OF SCIENCES HYPERTROPHIC CARD After 40 years of developing and utilizing assays to understand the James Spudich, Douglass M. and Nola Leishman Professor of molecular basis of energy transduction by the myosin family of Cardiovascular Disease, is in the Department of Biochemistry at Stanford molecular motors, all members of my laboratory are now focused on University School of Medicine. He received his B.S. in chemistry from understanding the underlying biochemical and biophysical bases of the University of Illinois in 1963 and his Ph.D. in biochemistry from human hypertrophic (HCM) and dilated (DCM) cardiomyopathies. Stanford in 1968. He did postdoctoral work in genetics at Stanford and Our primary focus is on HCM since these mutations cause the heart in structural biology at the MRC Laboratory in Cambridge, England. -
TALE-Light Imaging Reveals Maternally Guided, H3k9me2/3-Independent Emergence of Functional Heterochromatin in Drosophila Embryos
Downloaded from genesdev.cshlp.org on September 29, 2021 - Published by Cold Spring Harbor Laboratory Press TALE-light imaging reveals maternally guided, H3K9me2/3-independent emergence of functional heterochromatin in Drosophila embryos Kai Yuan and Patrick H. O’Farrell Department of Biochemistry, University of California at San Francisco, San Francisco, California 94158, USA Metazoans start embryogenesis with a relatively naïve genome. The transcriptionally inert, late-replicating het- erochromatic regions, including the constitutive heterochromatin on repetitive sequences near centromeres and telomeres, need to be re-established during development. To explore the events initiating heterochromatin forma- tion and examine their temporal control, sequence specificity, and immediate regulatory consequence, we estab- lished a live imaging approach that enabled visualization of steps in heterochromatin emergence on specific satellite sequences during the mid-blastula transition (MBT) in Drosophila. Unexpectedly, only a subset of satellite se- quences, including the 359-base-pair (bp) repeat sequence, recruited HP1a at the MBT. The recruitment of HP1a to the 359-bp repeat was dependent on HP1a’s chromoshadow domain but not its chromodomain and was guided by maternally provided signals. HP1a recruitment to the 359-bp repeat was required for its programmed shift to later replication, and ectopic recruitment of HP1a was sufficient to delay replication timing of a different repeat. Our results reveal that emergence of constitutive heterochromatin follows a stereotyped developmental program in which different repetitive sequences use distinct interactions and independent pathways to arrive at a heterochro- matic state. This differential emergence of heterochromatin on various repetitive sequences changes their replica- tion order and remodels the DNA replication schedule during embryonic development. -
Drosophila Ribosomal Proteins Are Associated with Linker Histone H1 and Suppress Gene Transcription
Downloaded from genesdev.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press Drosophila ribosomal proteins are associated with linker histone H1 and suppress gene transcription Jian-Quan Ni,1,3 Lu-Ping Liu,1,3 Daniel Hess,1 Jens Rietdorf,1 and Fang-Lin Sun1,2,4 1Friedrich Miescher Institute for Biomedical Research, Basel CH-4058, Switzerland; 2Institute of Epigenetics and Cancer Research, School of Medicine, Tsinghua University, Beijing 100080, China The dynamics and function of ribosomal proteins in the cell nucleus remain enigmatic. Here we provide evidence that specific components of Drosophila melanogaster ribosomes copurify with linker histone H1. Using various experimental approaches, we demonstrate that this association of nuclear ribosomal proteins with histone H1 is specific, and that colocalization occurs on condensed chromatin in vivo. Chromatin immunoprecipitation analysis confirmed that specific ribosomal proteins are associated with chromatin in a histone H1-dependent manner. Overexpression of either histone H1 or ribosomal protein L22 in Drosophila cells resulted in global suppression of the same set of genes, while depletion of H1 and L22 caused up-regulation of tested genes, suggesting that H1 and ribosomal proteins are essential for transcriptional gene repression. Overall, this study provides evidence for a previously undefined link between ribosomal proteins and chromatin, and suggests a role for this association in transcriptional regulation in higher eukaryotes. [Keywords: Ribosomal protein; L22; histone H1; chromatin; transcription] Supplemental material is available at http://www.genesdev.org. Received September 25, 2005; revised version accepted May 8, 2006. Transcription and translation in eukaryotes are generally discrete nuclear sites was sensitive to inhibitors of both believed to take place within two spatially separated cel- transcription and translation, arguing that the two pro- lular compartments. -
By Krista Conger It Was Around 3 P.M. on a Thursday Last October When James Spudich and Suzanne Pfeffer Poked Their Heads Into T
BECKMAN CENTER FOR MOLECULAR AND GENETIC MEDICINE PG. 6 BECKMAN RESEARCHERS MAP INTRICACIES OF LUNG CANCER IN ONE OF THEIR OWN By Krista Conger Spudich had no way of knowing it, but the meeting he’d interrupted between Krasnow and assistant professor of pediatrics Christin Kuo, MD, had been It was around 3 p.m. on a Thursday last October when called to discuss how Krasnow could broaden his James Spudich and Suzanne Pfeffer poked their heads research, which he had been conducting mainly in into the office of Mark Krasnow, on the fourth floor of mice and small primates called mouse lemurs, to the Beckman Center for Molecular and Genetic include human cancers. But the logistical and legal Medicine, interrupting a meeting he was having with hoops that would need to be cleared prior to any a colleague. kind of human-based research were daunting, and Krasnow knew it would likely take months to obtain “Can we talk to you for a minute?” Pfeffer said. the necessary approvals. Impromptu conversations were nothing new for the Kuo, a specialist in pulmonary medicine, had been three. The longtime colleagues and friends have a lot to working with Stephen Quake, PhD, professor of talk about. Spudich, PhD, whose research focuses on bioengineering and of applied physics and co-president understanding the molecular forces that drive muscle of the Chan Zuckerberg Biohub, and postdoctoral contractions, earned a doctoral degree from Stanford scholar Spyros Darmanis, PhD, a group leader at the in 1968 under Arthur Kornberg, MD, a founding Biohub, to study the development and function of lung member of the Department of Biochemistry. -
Introduction to the Genomics Education Partnership and Collaborative Genomics Research in Drosophila
Tested Studies for Laboratory Teaching Proceedings of the Association for Biology Laboratory Education Vol. 34, 135-165, 2013 Introduction to the Genomics Education Partnership and Collaborative Genomics Research in Drosophila Julia A. Emerson1, S. Catherine Silver Key2, Consuelo J. Alvarez3, Stephanie Mel4, Gerard P. McNeil5, Kenneth J. Saville6, Wilson Leung7, Christopher D. Shaffer7 and Sarah C. R. Elgin7 1Amherst College, Department of Biology, P.O. Box 5000, Amherst MA 01002 USA 2North Carolina Central University, Department of Biology, 2246 MTSB, Durham NC 27701 USA 3Longwood University, Department of Biological and Environmental Sciences, 201 High St., Farmville VA 23909 USA 4University of California San Diego, Division of Biological Sciences, 9500 Gilman Drive 0355, La Jolla CA 92093 USA 5York College/CUNY, Department of Biology, 94-20 Guy R. Brewer Blvd., Jamaica NY 11451 USA 6 Albion College, Biology Department, 611 E. Porter St., Albion MI 49224 USA 7Washington University, Department of Biology, Campus Box 1137, One Brookings Dr., St. Louis MO 3130 USA ([email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected]) The GEP, a group of faculty from over 90 primarily undergraduate institutions, is using comparative genomics to engage students in research within regular courses. Using a versatile curriculum, GEP undergraduates undertake projects to improve draft genomic sequences and/or participate in the annotation of these improved sequences. An additional goal of the annotation curriculum is for students to gain a sophisticated understanding of eukaryotic gene structure. Students do so by carefully mapping the protein-coding regions of putative genes from recently- sequenced Drosophila species. -
Workshop Notebook
GCAT WORKSHOPS 2009 NSF SUPPORTED DNA MICROARRAY WORKSHOPS FOR HISTORICALLY BLACK COLLEGES AND UNIVERSITIES, HISPANIC-SERVING INSTITUTIONS AND TRIBAL COLLEGES FACULTY JULY 5 - July 11, 2009 HOST INSTITUTION: MOREHOUSE COLLEGE BIOLOGY DEPARTMENT; ATLANTA, GA INSTRUCTORS Consuelo Alvarez, Malcolm Campbell, Todd Eckdahl, Edison Fowlks, Charles Hauser, Laurie Heyer, and Anne Rosenwald Genome Consortium for Active Teaching (GCAT) NSF GRANT DBI-0520908 Awarded to Hampton University PI: Edison R. Fowlks; Co-PIs: Mary Lee Ledbetter and Anne Rosenwald and NSF GRANT DBI-0520908 Awarded to Davidson College PI: Malcolm Campbell; Co-PIs: Laurie Heyer and Todd Eckdahl By the DIRECTORATE FOR BIOLOGICAL SCIENCES NATIONAL SCIENCE FOUNDATION DR. SALLY O’CONNOR, MANAGERING PROGRAM OFFICER NSF-SPONSORED GCAT DNA MICROARRAY SUMMER WORKSHOPS FOR UNDERGRADUATE HBCU, HSI AND TRIBAL COLLEGE FACULTY TAB I. Origin of this NSF Workshop and History of GCAT 1 II. Workshop Participants and Instructors 2 III. Workshop Schedule of Dry and Wet Lab Activities 3 IV. Assessment Material 4 A. Pre-Test B. Post-Test V. Wet Lab Activities 6 A. Culturing Yeast B. Isolating RNA C. 3DNA Protocol VI. Dry Lab Activities 7 A. Magic Tool User Guide B. Magic Tool: Exploring Diauxic Shift Microarray Data C. Exploring Correlations VII. Sampling of Microarray Articles 7 VIII. Notes, Special Instructions & Directions 8 ORIGIN OF THIS NSF WORKSHOP At a curriculum gathering during summer of 2004, Drs. Fowlks and Campbell presented their perspectives on the undergraduate educational implications of the NIH Roadmap. Fowlks addressed curricular changes in undergraduate biology, as well as recruitment and training for underrepresented populations. Campbell provided an overview of activities by GCAT, a non-profit educational consortium, to bring functional genomics methods into undergraduate courses and independent student research. -
Curriculum Vitae James A. Spudich, Ph.D
Curriculum Vitae James A. Spudich, Ph.D. Douglass M. and Nola Leishman Professor of Cardiovascular Disease Department of Biochemistry Stanford University School of Medicine EDUCATION 1963 University of Illinois, Chemistry Dept of Biochemistry B.S. 1968 Stanford University, Biochemistry Dept of Biochemistry Ph.D. 1968-1969 Stanford University, Molecular Genetics Dept of Biological Sciences Postdoctoral 1969-1971 Cambridge University, Structural Biology MRC Lab of Mol Biol Postdoctoral PROFESSIONAL EXPERIENCE 2012 Co-Founder, MyoKardia, Inc. 2011-present Adjunct Professor, Institute of Stem Cell Biology and Regenerative Medicine (inStem), Bangalore, India 2005-present Adjunct Professor, National Centre for Biological Sciences (NCBS) of the Tata Institute of Fundamental Research (TIFR), Bangalore, India; and TIFR, Bombay, India 1998-2002 Co-Founder and first Director, Interdisciplinary Program in Bioengineering, Biomedicine and Biosciences – Bio-X, Stanford University 1998 Co-Founder, Cytokinetics, Inc. 1992-present Professor, Dept of Biochemistry, Stanford University School of Medicine (Chairman from 1994-1998) 1989-2011 Professor, Dept of Developmental Biology, Stanford University School of Medicine 1977-1992 Professor, Dept of Structural Biology, Stanford University School of Medicine (Chairman from 1979-1984) 1976-1977 Professor, Dept of Biochemistry and Biophysics, UCSF, San Francisco 1974-1976 Associate Professor, Dept of Biochemistry and Biophysics, UCSF, San Francisco 1971-1974 Assistant Professor, Dept of Biochemistry and Biophysics, -
Michael Sheetz, James Spudich, and Ronald Vale Receive the 2012 Albert Lasker Basic Medical Research Award
Molecules in motion: Michael Sheetz, James Spudich, and Ronald Vale receive the 2012 Albert Lasker Basic Medical Research Award Sarah Jackson J Clin Invest. 2012;122(10):3374-3377. https://doi.org/10.1172/JCI66361. News The Albert and Mary Lasker Foundation honors three pioneers in the field of molecular motor proteins, Michael Sheetz, James Spudich, and Ronald Vale (Figure 1), for their contribution to uncovering how these proteins catalyze movement. Sheetz and Spudich developed the first biochemical assay to reconstitute myosin motor activity and showed that myosin and ATP alone were sufficient to direct transport along actin filaments. Fueled by this discovery, Vale and Sheetz examined transport in giant squid axon extracts and discovered a new motor, kinesin, that moves along microtubules. Cumulatively, their work opened the door for understanding how motor proteins drive transport of a host of molecules involved in numerous cellular processes, including cell polarity, cell division, cellular movement, and signal transduction. Muscle movement Much of the early research on cellular movement focused on understanding muscle contraction. Filaments of actin and myosin form a banding pattern in muscle that is readily visible by light microscopy, with a characteristic striated appearance. The functional unit of muscle, termed a sarcomere, repeats at regular intervals along the muscle fibers. Measurements from contracted, resting, and stretched muscle using high-resolution light microscopy supported the notion that contraction was due to the sliding of actin and myosin filaments (1, 2). In 1969, preeminent British scientist Hugh Huxley proposed that crossbridges that connect actin and myosin generate the sliding […] Find the latest version: https://jci.me/66361/pdf News Molecules in motion: Michael Sheetz, James Spudich, and Ronald Vale receive the 2012 Albert Lasker Basic Medical Research Award The Albert and Mary Lasker Founda- James Spudich joined the Huxley labora- motility along actin. -
Claire Barrett, Greg Creacy, Charles Hauser
Charles R. Hauser Associate Professor of Bioinformatics St. Edward’s University Voice: (512) 233.1671 Bioinformatics Program Fax. (512) 448.8764 Austin TX 78704 e-mail: [email protected] Professional Preparation University of Texas, Austin, TX 1972-1977 Botany University of Houston, TX 1984-1989 PhD. Biochemistry and Biophysics Duke University 1989-1995 Post-doctoral fellowship Appointments 2008 - Associate Professor Bioinformatics, St. Edward’s University 2004 - 2008 Assistant Professor Bioinformatics, St. Edward’s University 2000 - 2004 Research Scientist: Duke University, Durham NC 1995 - 2000 Research Associate: Duke University, Durham, NC 1989 - 1995 Postdoctoral Associate: Duke University, Durham, NC 1984 - 1989 Graduate Student/Researcher: University of Houston Awards and Honors 2016 St. Edward’s School of Natural Sciences Distinguished Research Award, 2015-16. 1990 NIH Postdoctoral Fellowship. Department of Botany, Duke University, Durham, NC. 1989 Hargitt Fellowship. Department of Zoology, Duke University, Durham, NC. Grants 2016 Howard Hughes Medical Institute (USE), Inclusive Excellence Grants 2017 Pre-proposal. Research L Kopec (PI), Charles R Hauser (collaborator) 2016 NSF, IUSE, “Integrating Bioinformatics into the Life Sciences—Phase 3” Mark Pauley (U. Nebraska, Omaha), Vince Buonaccorsi (Juniata College), Anya Goodman (California Polytechnic State University), Anne Rosenwald (Georgetown University) and Bill Tapprich (University of Nebraska at Omaha) Co-PIs. Charles Hauser (Advisory Committee Member). 2015 USDA, The Agricultural-STEM Pipeline: Progressive Experiential Learning Linking Three HSI Academic Tiers (Ag-STEM), 3 year ($275,000), Drs. O’Leary (PI), Quinn, Hauser, Deaton (co-PIs) 2015 NIH, “A Genome Browser On Ramp to Engage Biologists with Big Data”, Dr. Sarah Elgin, Washington University St. Louis (PI), Hauser (collaborator). -
Finding Genes in a New Fly Genome: Teaching About Genes/Genomes Via Bioinformatics Research
Finding Genes in a New Fly Genome: Teaching about Genes/Genomes via Bioinformatics Research Sarah Elgin, Anya Goodman, Wilson Leung Eric Tsoi, Charlene Emerson, David Carranza January 2012 Workshop Goals • Introduce Genomics Education Partnership • Hands-on practice with genome annotation • Discussion of curriculum options 3-week module ( ~10 hr: 1 lecture, 3X3 lab) 5-week – add a more difficult project 10-week - real research! • Scientific background on Drosophila genome • http://gep.wustl.edu [email protected] • Next workshops: June 24-26, August 19-22 (HHMI supported) Genomics Education Partnership (GEP) Goal: to engage students in a genomics research project, via a research-oriented AY lab course. Work organized through the GEP website. Our GEP research goal: Use comparative genomics to learn more about heterochromatic domains, analyzing the dot chromosomes and a control euchromatic region of Drosophila genomes Status Reference Completed Annotation Sequence Improvement New Project FlyBase: http://flybase.org I hear and I forget. I see and I remember. I do and I understand Confucius The scientific method allows ordinary people to do extraordinary things. Francis Bacon Genomics provides terrific opportunities to engage undergraduates in research! Strategy: divide and conquer! The D. mojavensis dot chromosome • Students completed 68 projects covering 1.7 Mb closing 26/28 gaps, adding ~15,000 bp and improving ~5000 bp. • Each project finished and annotated (all isoforms) twice; reconciliation for quality control done at Wash U Fosmid -
Heterochromatin-Associated Interactions of Drosophila Hp1a with Dadd1, HIPP1, and Repetitive Rnas
Downloaded from genesdev.cshlp.org on October 3, 2021 - Published by Cold Spring Harbor Laboratory Press Heterochromatin-associated interactions of Drosophila HP1a with dADD1, HIPP1, and repetitive RNAs Artyom A. Alekseyenko,1,2 Andrey A. Gorchakov,1,2,3 Barry M. Zee,1,2 Stephen M. Fuchs,4 Peter V. Kharchenko,5,6,7 and Mitzi I. Kuroda1,2,7 1Division of Genetics, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA; 2Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA; 3Institute of Molecular and Cellular Biology, Novosibirsk 630090, Russia; 4Department of Biology, Tufts University, Medford, Massachusetts 02155, USA; 5Center for Biomedical Informatics, Harvard Medical School, Boston, Massachusetts 02115, USA; 6Hematology/Oncology Program, Children’s Hospital, Boston, Massachusetts 02115, USA Heterochromatin protein 1 (HP1a) has conserved roles in gene silencing and heterochromatin and is also implicated in transcription, DNA replication, and repair. Here we identify chromatin-associated protein and RNA interactions of HP1a by BioTAP-XL mass spectrometry and sequencing from Drosophila S2 cells, embryos, larvae, and adults. Our results reveal an extensive list of known and novel HP1a-interacting proteins, of which we selected three for validation. A strong novel interactor, dADD1 (Drosophila ADD1) (CG8290), is highly enriched in heterochromatin, harbors an ADD domain similar to human ATRX, displays selective binding to H3K9me2 and H3K9me3, and is a classic genetic suppressor of position-effect variegation. Unexpectedly, a second hit, HIPP1 (HP1 and insulator partner protein-1) (CG3680), is strongly connected to CP190-related complexes localized at putative insulator sequences throughout the genome in addition to its colocalization with HP1a in heterochro- matin.