Characterization of E Coli Hfq Structure and Its Rna Binding Properties
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RNA-Binding Protein Network Alteration Causes Aberrant Axon
bioRxiv preprint doi: https://doi.org/10.1101/2020.08.26.268631; this version posted August 26, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.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-NC-ND 4.0 International license. 1 RNA-binding protein network alteration causes aberrant axon 2 branching and growth phenotypes in FUS ALS mutant motoneurons 3 4 Maria Giovanna Garone1, Nicol Birsa2,3, Maria Rosito4, Federico Salaris1,4, Michela Mochi1, Valeria 5 de Turris4, Remya R. Nair5, Thomas J. Cunningham5, Elizabeth M. C. Fisher2, Pietro Fratta2 and 6 Alessandro Rosa1,4,6,* 7 8 1. Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le 9 A. Moro 5, 00185 Rome, Italy 10 2. UCL Queen Square Institute of Neurology, University College London, London, WC1N 3BG, 11 UK 12 3. UK Dementia Research Institute, University College London, London, WC1E 6BT, UK 13 4. Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, 00161 14 Rome, Italy 15 5. MRC Harwell Institute, Oxfordshire, OX11 0RD, UK 16 6. Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci Bolognetti, Department of 17 Biology and Biotechnology Charles Darwin, Sapienza University of Rome, Viale Regina Elena 18 291, 00161 Rome, Italy 19 20 * Corresponding author: [email protected]; Tel: +39-0649255218 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.08.26.268631; this version posted August 26, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. -
Compaction and Condensation of DNA Mediated by the C-Terminal Domain Of
i “CTR-NAR” — 2017/5/3 — 11:51 — page 1 — #1 i i i Published online 03/05/2017 Nucleic Acids Research, 2017, Vol. 00, No. 00 1–10 doi:10.1093/nar/gkn000 Compaction and condensation of DNA mediated by the C-terminal domain of Hfq 1, 2, 1,3 4 Antoine Malabirade †, Kai Jiang †, Krzysztof Kubiak , Alvaro Diaz-Mendoza , Fan Liu 2, Jeroen A. van Kan 2, Jean-François Berret 4, 1,4, 2, Véronique Arluison ú, and Johan R.C. van der Maarel ú 1Laboratoire Léon Brillouin, CEA, CNRS, Université Paris Saclay, 91191 Gif-sur-Yvette, France; 2Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore 117542; 3Department of Molecular Biology, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk, Poland; 4Université Paris Diderot, Sorbonne Paris Cité, 75013 Paris, France. ABSTRACT it is usually needed to mediate post-transcriptional stress-response with small noncoding RNA (sRNA). This Hfq is a bacterial protein that is involved in several regulatory mechanism is based on the hybridisation of aspects of nucleic acids metabolism. It has been sRNA to its target mRNA. Accordingly, the majority described as one of the nucleoid associated proteins of known sRNA interacts nearby the ribosome binding shaping the bacterial chromosome, although it is site to prevent initiation of translation and thus favours better known to influence translation and turnover mRNA decay (4, 5, 6). Few studies have shed light of cellular RNAs. Here, we explore the role of on the importance of Hfq in DNA metabolism. It has Escherichia coli Hfq’s C-terminal domain in the been suggested that Hfq affects plasmid replication and compaction of double stranded DNA. -
Emerging Roles for 3 Utrs in Neurons
International Journal of Molecular Sciences Review 0 Emerging Roles for 3 UTRs in Neurons Bongmin Bae and Pedro Miura * Department of Biology, University of Nevada, Reno, NV 89557, USA; [email protected] * Correspondence: [email protected] Received: 8 April 2020; Accepted: 9 May 2020; Published: 12 May 2020 Abstract: The 30 untranslated regions (30 UTRs) of mRNAs serve as hubs for post-transcriptional control as the targets of microRNAs (miRNAs) and RNA-binding proteins (RBPs). Sequences in 30 UTRs confer alterations in mRNA stability, direct mRNA localization to subcellular regions, and impart translational control. Thousands of mRNAs are localized to subcellular compartments in neurons—including axons, dendrites, and synapses—where they are thought to undergo local translation. Despite an established role for 30 UTR sequences in imparting mRNA localization in neurons, the specific RNA sequences and structural features at play remain poorly understood. The nervous system selectively expresses longer 30 UTR isoforms via alternative polyadenylation (APA). The regulation of APA in neurons and the neuronal functions of longer 30 UTR mRNA isoforms are starting to be uncovered. Surprising roles for 30 UTRs are emerging beyond the regulation of protein synthesis and include roles as RBP delivery scaffolds and regulators of alternative splicing. Evidence is also emerging that 30 UTRs can be cleaved, leading to stable, isolated 30 UTR fragments which are of unknown function. Mutations in 30 UTRs are implicated in several neurological disorders—more studies are needed to uncover how these mutations impact gene regulation and what is their relationship to disease severity. Keywords: 30 UTR; alternative polyadenylation; local translation; RNA-binding protein; RNA-sequencing; post-transcriptional regulation 1. -
Producing Hfq/Sm Proteins and Srnas for Structural and Biophysical Studies of Ribonucleoprotein Assembly
bioRxiv preprint doi: https://doi.org/10.1101/150672; this version posted June 16, 2017. 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. Producing Hfq/Sm Proteins and sRNAs for Structural and Biophysical Studies of Ribonucleoprotein Assembly Kimberly A. Stanek* and Cameron Mura* Author affiliations & correspondence: Department of Chemistry; University of Virginia; Charlottesville, VA 22904 USA *Correspondence can be addressed to KS ([email protected]) or CM ([email protected]). Document information: Last modified: 15 June 2017 Running title: Producing Hfq•RNA Complexes for Structural Studies Abbreviations: 3D, three-dimensional; AU, asymmetric unit; CV, column volume; DEPC, diethyl pyrocarbonate; HDV, hepatitis δ virus; IMAC, immobilized metal affinity chroma- tography; MW, molecular weight; MWCO, molecular weight cut-off; nt, nucleotide; PDB, Protein Data Bank; RNP, ribonucleoprotein; RT, room temperature Additional notes: The main text is accompanied by 2 figures and 1 table. Journal format: Methods in Molecular Biology (Springer Protocols series); this volume is entitled "Bacterial Regulatory RNA: Methods and Protocols"; an author guide is linked at http://www.springer.com/series/7651 bioRxiv preprint doi: https://doi.org/10.1101/150672; this version posted June 16, 2017. 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 Hfq is a bacterial RNA-binding protein that plays key roles in the post–transcriptional regulation of gene expression. Like other Sm proteins, Hfq assembles into toroidal discs that bind RNAs with varying affin- ities and degrees of sequence specificity. -
New Insights Into Functional Roles of the Polypyrimidine Tract-Binding Protein
Int. J. Mol. Sci. 2013, 14, 22906-22932; doi:10.3390/ijms141122906 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review New Insights into Functional Roles of the Polypyrimidine Tract-Binding Protein Maria Grazia Romanelli *, Erica Diani and Patricia Marie-Jeanne Lievens Department of Life and Reproduction Sciences, Section of Biology and Genetics, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy; E-Mails: [email protected] (E.D.); [email protected] (P.M.-J.L.) * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +39-045-8027182; Fax: +39-045-8027180. Received: 22 September 2013; in revised form: 13 November 2013 / Accepted: 13 November 2013 / Published: 20 November 2013 Abstract: Polypyrimidine Tract Binding Protein (PTB) is an intensely studied RNA binding protein involved in several post-transcriptional regulatory events of gene expression. Initially described as a pre-mRNA splicing regulator, PTB is now widely accepted as a multifunctional protein shuttling between nucleus and cytoplasm. Accordingly, PTB can interact with selected RNA targets, structural elements and proteins. There is increasing evidence that PTB and its paralog PTBP2 play a major role as repressors of alternatively spliced exons, whose transcription is tissue-regulated. In addition to alternative splicing, PTB is involved in almost all steps of mRNA metabolism, including polyadenylation, mRNA stability and initiation of protein translation. Furthermore, it is well established that PTB recruitment in internal ribosome entry site (IRES) activates the translation of picornaviral and cellular proteins. Detailed studies of the structural properties of PTB have contributed to our understanding of the mechanism of RNA binding by RNA Recognition Motif (RRM) domains. -
And Plasmid-Borne Resistance of Escherichia Coli Hfq Mutants to High Concentrations of Various Antibiotics
International Journal of Molecular Sciences Article Differential Chromosome- and Plasmid-Borne Resistance of Escherichia coli hfq Mutants to High Concentrations of Various Antibiotics Lidia Gaffke † , Krzysztof Kubiak †, Zuzanna Cyske and Grzegorz W˛egrzyn* Department of Molecular Biology, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk, Poland; [email protected] (L.G.); [email protected] (K.K.); [email protected] (Z.C.) * Correspondence: [email protected]; Tel.: +48-58-523-6024 † These authors contributed equally to this work. Abstract: The Hfq protein is a bacterial RNA chaperone, involved in many molecular interactions, including control of actions of various small RNA regulatory molecules. We found that the presence of Hfq was required for survival of plasmid-containing Escherichia coli cells against high concentrations of chloramphenicol (plasmid p27cmr), tetracycline (pSC101, pBR322) and ampicillin (pBR322), as hfq+ strains were more resistant to these antibiotics than the hfq-null mutant. In striking contrast, production of Hfq resulted in low resistance to high concentrations of kanamycin when the antibiotic- resistance marker was chromosome-borne, with deletion of hfq resulting in increasing bacterial survival. These results were observed both in solid and liquid medium, suggesting that antibiotic resistance is an intrinsic feature of these strains rather than a consequence of adaptation. Despite its major role as RNA chaperone, which also affects mRNA stability, Hfq was not found to significantly affect kan and tet mRNAs turnover. Nevertheless, kan mRNA steady-state levels were higher in Citation: Gaffke, L.; Kubiak, K.; the hfq-null mutant compared to the hfq+ strain, suggesting that Hfq can act as a repressor of kan Cyske, Z.; W˛egrzyn,G. -
Comparative Interactomics Analysis of Different ALS-Associated Proteins
Acta Neuropathol (2016) 132:175–196 DOI 10.1007/s00401-016-1575-8 ORIGINAL PAPER Comparative interactomics analysis of different ALS‑associated proteins identifies converging molecular pathways Anna M. Blokhuis1 · Max Koppers1,2 · Ewout J. N. Groen1,2,9 · Dianne M. A. van den Heuvel1 · Stefano Dini Modigliani4 · Jasper J. Anink5,6 · Katsumi Fumoto1,10 · Femke van Diggelen1 · Anne Snelting1 · Peter Sodaar2 · Bert M. Verheijen1,2 · Jeroen A. A. Demmers7 · Jan H. Veldink2 · Eleonora Aronica5,6 · Irene Bozzoni3 · Jeroen den Hertog8 · Leonard H. van den Berg2 · R. Jeroen Pasterkamp1 Received: 22 January 2016 / Revised: 14 April 2016 / Accepted: 15 April 2016 / Published online: 10 May 2016 © The Author(s) 2016. This article is published with open access at Springerlink.com Abstract Amyotrophic lateral sclerosis (ALS) is a dev- OPTN and UBQLN2, in which mutations caused loss or astating neurological disease with no effective treatment gain of protein interactions. Several of the identified inter- available. An increasing number of genetic causes of ALS actomes showed a high degree of overlap: shared binding are being identified, but how these genetic defects lead to partners of ATXN2, FUS and TDP-43 had roles in RNA motor neuron degeneration and to which extent they affect metabolism; OPTN- and UBQLN2-interacting proteins common cellular pathways remains incompletely under- were related to protein degradation and protein transport, stood. To address these questions, we performed an inter- and C9orf72 interactors function in mitochondria. To con- actomic analysis to identify binding partners of wild-type firm that this overlap is important for ALS pathogenesis, (WT) and ALS-associated mutant versions of ATXN2, we studied fragile X mental retardation protein (FMRP), C9orf72, FUS, OPTN, TDP-43 and UBQLN2 in neuronal one of the common interactors of ATXN2, FUS and TDP- cells. -
Review Article Proteins with RNA Chaperone Activity: a World of Diverse Proteins with a Common Task—Impediment of RNA Misfolding
Hindawi Publishing Corporation Biochemistry Research International Volume 2011, Article ID 532908, 11 pages doi:10.1155/2011/532908 Review Article Proteins with RNA Chaperone Activity: A World of Diverse Proteins with a Common Task—Impediment of RNA Misfolding Katharina Semrad Department of Biochemistry and Cell Biology, Max F. Perutz Laboratories, University of Vienna, Dr. Bohrgasse 9/5, 1030 Vienna, Austria Correspondence should be addressed to Katharina Semrad, [email protected] Received 19 July 2010; Revised 12 November 2010; Accepted 19 November 2010 Academic Editor: Andrei Surguchov Copyright © 2011 Katharina Semrad. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Proteins with RNA chaperone activity are ubiquitous proteins that play important roles in cellular mechanisms. They prevent RNA from misfolding by loosening misfolded structures without ATP consumption. RNA chaperone activity is studied in vitro and in vivo using oligonucleotide- or ribozyme-based assays. Due to their functional as well as structural diversity, a common chaperoning mechanism or universal motif has not yet been identified. A growing database of proteins with RNA chaperone activity has been established based on evaluation of chaperone activity via the described assays. Although the exact mechanism is not yet understood, it is more and more believed that disordered regions within proteins play an important role. This possible mechanism and which proteins were found to possess RNA chaperone activity are discussed here. 1. Introduction In the cellular environment, RNA molecules do not appear as “naked” nucleic acids but always are found in Among all biological macromolecules, RNAs represent one conjunction with proteins. -
Intermolecular Base Stacking Mediates RNA-RNA Interaction in a Crystal
www.nature.com/scientificreports OPEN Intermolecular base stacking mediates RNA-RNA interaction in a crystal structure of the RNA Received: 12 April 2017 Accepted: 2 August 2017 chaperone Hfq Published: xx xx xxxx Eike C. Schulz1,2,5, Markus Seiler1,6, Cecilia Zuliani1, Franka Voigt1,7, Vladimir Rybin3, Vivian Pogenberg 2, Norbert Mücke4, Matthias Wilmanns2, Toby J. Gibson1 & Orsolya Barabas1 The RNA-chaperone Hfq catalyses the annealing of bacterial small RNAs (sRNAs) with target mRNAs to regulate gene expression in response to environmental stimuli. Hfq acts on a diverse set of sRNA- mRNA pairs using a variety of diferent molecular mechanisms. Here, we present an unusual crystal structure showing two Hfq-RNA complexes interacting via their bound RNA molecules. The structure contains two Hfq6:A18 RNA assemblies positioned face-to-face, with the RNA molecules turned towards each other and connected via interdigitating base stacking interactions at the center. Biochemical data further confrm the observed interaction, and indicate that RNA-mediated contacts occur between Hfq-RNA complexes with various (ARN)X motif containing RNA sequences in vitro, including the stress response regulator OxyS and its target, flA. A systematic computational survey also shows that phylogenetically conserved (ARN)X motifs are present in a subset of sRNAs, some of which share similar modular architectures. We hypothesise that Hfq can co-opt RNA-RNA base stacking, an unanticipated structural trick, to promote the interaction of (ARN)X motif containing sRNAs with target mRNAs on a “speed-dating” fashion, thereby supporting their regulatory function. Non-coding RNAs play key roles in regulating gene expression in all domains of life. -
Back Matter (PDF)
JOBNAME: RNA 12#12 2006 PAGE: 1 OUTPUT: November 10 14:27:49 2006 csh/RNA/125784/reviewers-index RNA: Reviewers for Volume 12, 2006 The editors wish to thank the following individuals whose efforts in reviewing papers for RNA in the past year are greatly appreciated. Juan Alfonzo Donald Burke Fritz Eckstein Carol Greider Frederic Allain John Burke Martin Egli Sam Griffiths-Jones Emily Allen Samuel Butcher Sherif Abou Elela Claudio Gualerzi Sidney Altman Ronald Emeson Alexander Gultyaev Victor Ambros Gail Emilsson Samuel Gunderson Mark Caprara James Anderson Luis Enjuanes Christine Guthrie Paul Anderson Massimo Caputi Anne Ephrussi Raul Andino James Carrington Jay Evans Charles Carter Gordon Hager Mohammed Ararzguioui Eduardo Eyras Paul Hagerman Manuel Ares Richard Carthew Jamie Cate Stephen Hajduk Jean Armengaud Dan Fabris Jean Cavarelli Kathleen Hall Brandon Ason Philip Farabaugh Guillaume Chanfreau Michelle Hamm Gil Ast Jean Feagin Tien-Hsien Chang Scott Hammond Pascal Auffinger Martha Fedor Lawrence Chasin Maureen Hanson Johanna Avis Yuriy Fedorov Chang-Zheng Chen Eoghan Harrington Juli Feigon Eric Christian Michael Harris James Fickett Kristian Baker Christine Clayton Roland Hartmann Carol Fierke Alice Barkan Peter Clote Stephen Harvey Susan Baserga Witold Filipowicz Jeffery Coller Michelle Hastings Brenda Bass Andrew Fire Kathleen Collins Christopher Hayes Christoph Flamm David Bartel Richard Collins Christopher Hellen William Folk Robert Batey Elena Conti Matthias Hentze Maurille Fournier Peter Becker Howard Cooke Thomas Herdegen -
This Thesis Has Been Submitted in Fulfilment of the Requirements for a Postgraduate Degree (E.G
This thesis has been submitted in fulfilment of the requirements for a postgraduate degree (e.g. PhD, MPhil, DClinPsychol) at the University of Edinburgh. Please note the following terms and conditions of use: • This work is protected by copyright and other intellectual property rights, which are retained by the thesis author, unless otherwise stated. • A copy can be downloaded for personal non-commercial research or study, without prior permission or charge. • This thesis cannot be reproduced or quoted extensively from without first obtaining permission in writing from the author. • The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the author. • When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given. Expression and subcellular localisation of poly(A)-binding proteins Hannah Burgess PhD The University of Edinburgh 2010 Abstract Poly(A)-binding proteins (PABPs) are important regulators of mRNA translation and stability. In mammals four cytoplasmic PABPs with a similar domain structure have been described - PABP1, tPABP, PABP4 and ePABP. The vast majority of research on PABP mechanism, function and sub-cellular localisation is however limited to PABP1 and little published work has explored the expression of PABP proteins. Here, I examine the tissue distribution of PABP1 and PABP4 in mouse and show that both proteins differ markedly in their expression at both the tissue and cellular level, contradicting the widespread perception that PABP1 is ubiquitously expressed. PABP4 is shown to be widely expressed though with an expression pattern distinct from PABP1, and thus may have a biological function in many tissues. -
Investigation of RNA Binding Proteins Regulated by Mtor
Investigation of RNA binding proteins regulated by mTOR Thesis submitted to the University of Leicester for the degree of Doctor of Philosophy Katherine Morris BSc (University of Leicester) March 2017 1 Investigation of RNA binding proteins regulated by mTOR Katherine Morris, MRC Toxicology Unit, University of Leicester, Leicester, LE1 9HN The mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase which plays a key role in the transduction of cellular energy signals, in order to coordinate and regulate a wide number of processes including cell growth and proliferation via control of protein synthesis and protein degradation. For a number of human diseases where mTOR signalling is dysregulated, including cancer, the clinical relevance of mTOR inhibitors is clear. However, understanding of the mechanisms by which mTOR controls gene expression is incomplete, with implications for adverse toxicological effects of mTOR inhibitors on clinical outcomes. mTOR has been shown to regulate 5’ TOP mRNA expression, though the exact mechanism remains unclear. It has been postulated that this may involve an intermediary factor such as an RNA binding protein, which acts downstream of mTOR signalling to bind and regulate translation or stability of specific messages. This thesis aimed to address this question through the use of whole cell RNA binding protein capture using oligo‐d(T) affinity isolation and subsequent proteomic analysis, and identify RNA binding proteins with differential binding activity following mTOR inhibition. Following validation of 4 identified mTOR‐dependent RNA binding proteins, characterisation of their specific functions with respect to growth and survival was conducted through depletion studies, identifying a promising candidate for further work; LARP1.