Contributors: Architecture et fonction des macromolécules biologiques (AFMB); Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE); Centro de Investigaciones Biológicas (CSIC); Consejo Superior de Investigaciones Científicas [Madrid] (CSIC); Institut de Recherche Saint-Louis - Hématologie Immunologie Oncologie (Département de recherche de l’UFR de médecine; ex- Institut Universitaire Hématologie-IUH) (IRSL); Université Paris Cité (UPCité); Génomes, biologie cellulaire et thérapeutiques (GenCellDi (U944 / UMR7212)); Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité); Institut de Biologie Intégrative de la Cellule (I2BC); Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS); the Spanish Ministry of Science / Agencia Estatal de Investigación (BFU2017-87397-P, RED2018-102467-T and PID2020-116722GB-I00 to C. F.-T.; PRE2018-087012 to A. P.-P.), intramural funding from CSIC (2020AEP152 and PIE-202120E047-Conexiones-Life to C. F.-T.), Fondation pour la Recherche Médicale (FRM-EQU202203014635 to P. L.; FRM-SPF20170938755 to A. A.-L.; FRM-ECO202206015504 to B. C.), and intramural funding from CNRS, Université de Paris and INSERM (to P. L.); ANR-17-CE11-0025,INstruc,Etude structurale du mécanisme d'intégration des rétrotransposons de levure dans les gènes transcrits par l'ARN Polymérase III(2017); Agence Nationale de la Recherche (France); Agencia Estatal de Investigación (España); Consejo Superior de Investigaciones Científicas (España); Fondation pour la Recherche Médicale; Centre National de la Recherche Scientifique (France); Université Paris Cité; Institut National de la Santé et de la Recherche Médicale (France); Nguyen, Phong Quoc; Huecas, Sonia; Asif-Laidin, Amna; Plaza-Pegueroles, Adrián; Capuzzi, Beatrice; Palmic, Noé; Acker, Joël; Reguera, Juan; Lesage, Pascale; Fernández-Tornero, Carlos
نبذة مختصرة : 13 p.-6 fig.
The yeast Ty1 retrotransposon integrates upstream of genes transcribed by RNA polymerase III (Pol III). Specificity of integration is mediated by an interaction between the Ty1 integrase (IN1) and Pol III, currently uncharacterized at the atomic level. We report cryo-EM structures of Pol III in complex with IN1, revealing a 16-residue segment at the IN1 C-terminus that contacts Pol III subunits AC40 and AC19, an interaction that we validate by in vivo mutational analysis. Binding to IN1 associates with allosteric changes in Pol III that may affect its transcriptional activity. The C-terminal domain of subunit C11, involved in RNA cleavage, inserts into the Pol III funnel pore, providing evidence for a two-metal mechanism during RNA cleavage. Additionally, ordering next to C11 of an N-terminal portion from subunit C53 may explain the connection between these subunits during termination and reinitiation. Deletion of the C53 N-terminal region leads to reduced chromatin association of Pol III and IN1, and a major fall in Ty1 integration events. Our data support a model in which IN1 binding induces a Pol III configuration that may favor its retention on chromatin, thereby improving the likelihood of Ty1 integration.
This work was supported by the Agence Nationale de la Recherche (ANR-17-CE11-0025 to P.L., J.A., J.R. and C.F.-T.), the Spanish Ministry of Science/Agencia Estatal de Investigación (BFU2017-87397-P, RED2018-102467-T and PID2020-116722GB-I00 to C.F.-T.; PRE2018-087012 to A.P.-P.), intramural funding from CSIC (2020AEP152 and PIE-202120E047-Conexiones-Life to C. F.-T.), Fondation pour la Recherche Médicale (FRM-EQU202203014635 to P.L.; FRM-SPF20170938755 to A.A.-L.; FRM-ECO202206015504 to B.C.), and intramural funding from CNRS, Université Paris Cité and INSERM (to P.L.).
_1.png)
Processing Request
No Comments.