Aspects moléculaires des hélicases de la famille de RecQ Hua Ren To cite this version: Hua Ren. Aspects moléculaires des hélicases de la famille de RecQ. Sciences du Vivant [q-bio]. École normale supérieure de Cachan - ENS Cachan, 2009. Français. tel-00448084 HAL Id: tel-00448084 https://tel.archives-ouvertes.fr/tel-00448084 Submitted on 18 Jan 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THESE DE DOCTORAT DE L’ECOLE NORMALE SUPERIEURE DE CACHAN Présentée par Madame Hua REN pour obtenir le grade de DOCTEUR DE L’ECOLE NORMALE SUPERIEURE DE CACHAN Domaine : SCIENCES DE LA VIE ET DE LA SANTE Sujet de la thèse: Aspects moléculaires des hélicases de la famille de RecQ Thèse présentée et soutenue à ECNU Chine le 28 / 09/ 2009 devant le jury composé de : M. Marc BOUDVILLAIN Président/Rapporteur M. Jie-min WENG Rapporteur M. Duan MA Examinateur Mme Min QIAN Co-Directrice de thèse M. Xu-guang XI Co-Directeur de thèse Nom du Laboratoire: LBPA ENS CACHAN/CNRS/UMR 8113 61, avenue du Président Wilson, 94235 CACHAN CEDEX (France) ACKNOWLEDGEMENT This thesis would not have been done possibly without the support of many people. Foremost, I would like to express my gratitude to my two co-supervisors, Prof. Min QIAN and Prof. Xu-guang XI. Thanks to Xu-guang XI for allowing me to join his team, sharing with me a lot of his expertise and research insight. He is extremely helpful and offering me invaluable assistance, support and guidance. I would like to thank Prof. Min QIAN for her expertise, kindness, patience and most of all, for her support. My great appreciation also goes to my thesis committee members: Prof. Marc BOUDVILLAIN, Prof. Jie-min WENG, Prof. Duan MA, Prof. Jia LI and Prof. Xiang-huo HE. Without their knowledgeable and considerable assistances, this thesis would not have been successful. I am greatly indebted to many teachers in the Ecole normale supérieure de Cachan (ENS Cachan), East China Normal University and Institute Curie of Orsay. Especially thanks to Prof. Christian AUCLAIR, the director of the laboratories and institutes of ENS Cachan; Prof. Jean-francois MOUSCADET, the director of Laboratory and Applied Genetic Pharmacology Laboratory (LBPA); Prof. Giuseppe Baldacci, the president of Institute Curie of Orsay; Mesdames Bogdana NEUVILLE and Yu-hua QIAN; Mr. Hai-sheng LI. And I am deeply grateful to the French government, the Chinese Scholarship Council and the Chinese Embassy in France for providing me the financial support during my PhD. Studies. I am tempted to individually thank all of the members in the same team, Mesdames Rong-bing GUO, Min ZHOU, Françoise CHAMINADE, Mounira AMOR-GUERET, Rosine Onclercq-Delic, Emmanuelle Delagoutte; Messieurs Pascal Rigolet, Philippe FOSSE, Nicolas BAZEILLE, Jie-lin LIU, Bo ZHANG, Da-peng ZHANG. It is never been overstated how much thanks I owes to all my friends who studied with me together in France. Thank you for your company which made my lives there enjoyable and beyond my expectation. Last but not least, I want to show how grateful I am with my family: my husband Wei NIU who has been standing beside me and encouraging me as always; my mother Wan- ying ZHANG whose love is boundless and my father Hui-bin REN who is my role model. They have always been supporting and stimulating me to do my best in all matters of life. To them I heartily dedicate this thesis. 2 TABLE OF CONTENTS SUMMARY IN CHINESE （中文摘要）········································································································· 7 SUMMARY IN FRENCH (RESUME) ············································································································ 11 SUMMARY ························································································································································· 13 ABREVIATIONS ··············································································································································· 15 CHAPTER 1 GENERAL INTRODUCTION ························································································· 18 1. DIFFERENT HELICASE FAMILIES ········································································································ 20 SUPERFAMILY 1 ·········································································································································· 21 SUPERFAMILY 2 ·········································································································································· 21 SUPERFAMILY 3 ·········································································································································· 22 SUPERFAMILY 4 ·········································································································································· 24 OTHER SUPERFAMILIES ······························································································································ 26 2. HELICASE-GENERAL PROPERTIES OF HELICASES AND MODE OF ACTION ······················· 27 2.1 BIOCHEMICAL PROPERTIES OF HELICASES ····························································································· 27 2.1.1 NTP binding and hydrolysis of NTP ······························································································· 27 2.1.2 Nucleic Acids binding ····················································································································· 27 2.1.3 Unwinding Activity and Mode of Action ························································································· 28 2.1.4 Annealing Activity ·························································································································· 31 2.2 THE MULTIPLE FUNCTIONS OF HELICASES ····························································································· 32 Helicase and DNA replication ················································································································· 32 Helicase and DNA repair ························································································································ 33 Helicase and RNA metabolism ················································································································ 33 3. THE RECQ FAMILY OF DNA HELICASES ··························································································· 34 3.1 GENERAL FEATURES AND STRUCTURAL ORGANISATION ······································································· 34 3. 2 THE ESCHERICHIA COLI RECQ ·············································································································· 39 3.2.1 Genetic Background ······················································································································· 39 3.2.2 Biochemical and structural Properties of E.coli RecQ ·································································· 39 3.2.3 Role in genomic stability ················································································································ 40 3.2.4 RecQ helicases from other microorganisms ··················································································· 43 3.3 YEAST RECQ FAMILY HELICASES ·········································································································· 43 3.3.1 Saccharomyces cerevisiae Sgs1 ······································································································ 43 3.3.1.1 Sgs1 Cellular Phenotype ············································································································· 43 3.3.1.2 Biochemical Properties of Sgs1 ······························································································· 44 3.3.1.3 Interaction Partners and Models for genome instability ························································· 44 3.3.2 Schizosaccharomyces pombe Rqh1 ································································································ 46 3.3.2.1 Biochemical properites ·························································································································· 46 3.3.2.2 Function in telomere metabolism ··········································································································· 46 3.4 RECQ HELICASES IN HUMAN ················································································································· 47 3.4.1 Human RECQ1 ······························································································································· 47 Human RECQl and genome stability ················································································································· 47 Biochemical properties ······································································································································