title: Yeast mitochondrial RNA polymerase primes mitochondrial DNA polymerase at origins of replication and promoter sequences.
abstract: Three proteins phylogenetically grouped with proteins from the T7 replisome localize to yeast mitochondria: DNA polymerase γ (Mip1), mitochondrial RNA polymerase (Rpo41), and a single-stranded binding protein (Rim1). Human and T7 bacteriophage RNA polymerases synthesize primers for their corresponding DNA polymerases. In contrast, DNA replication in yeast mitochondria is explained by two models: a transcription-dependent model in which Rpo41 primes Mip1 and a model in which double stranded breaks create free 3' OHs that are extended by Mip1. Herein we found that Rpo41 transcribes RNAs that can be extended by Mip1 on single and double-stranded DNA. In contrast to human mitochondrial RNA polymerase, which primes DNA polymerase γ using transcripts from the light-strand and heavy-strand origins of replication, Rpo41 primes Mip1 at replication origins and promoter sequences in vitro. Our results suggest that in ori1, short transcripts serve as primers, whereas in ori5 an RNA transcript longer than 29 nucleotides is used as primer.
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Sanchez-Sandoval
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Corina
authors.initials: C
authors.lastname: Diaz-Quezada
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Gilberto
authors.initials: G
authors.lastname: Velazquez
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Luis F
authors.initials: LF
authors.lastname: Arroyo-Navarro
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Norineli
authors.initials: N
authors.lastname: Almanza-Martinez
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Carlos H
authors.initials: CH
authors.lastname: Trasviña-Arenas
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, CP 36500 Irapuato, Guanajuato, Mexico. Electronic address: lgbrieba@langebio.cinvestav.mx.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Brieba
mesh_terms: D004256:DNA Polymerase I / Q000378:metabolism*; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000096:biosynthesis*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology* / Q000235:genetics; D024101:Mitochondrial Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D018741:Replication Origin; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D004272:DNA, Mitochondrial; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D012321:DNA-Directed RNA Polymerases; C487996:RPO41 protein, S cerevisiae; D004256:DNA Polymerase I; C068352:MIP1 protein, S cerevisiae
keywords: In vitro; RPO41; Replication; Yeast mitochondria