title: The thumb subdomain of yeast mitochondrial RNA polymerase is involved in processivity, transcript fidelity and mitochondrial transcription factor binding.
abstract: Single subunit RNA polymerases have evolved 2 mechanisms to synthesize long transcripts without falling off a DNA template: binding of nascent RNA and interactions with an RNA:DNA hybrid. Mitochondrial RNA polymerases share a common ancestor with T-odd bacteriophage single subunit RNA polymerases. Herein we characterized the role of the thumb subdomain of the yeast mtRNA polymerase gene (RPO41) in complex stability, processivity, and fidelity. We found that deletion and point mutants of the thumb subdomain of yeast mtRNA polymerase increase the synthesis of abortive transcripts and the probability that the polymerase will disengage from the template during the formation of the late initial transcription and elongation complexes. Mutations in the thumb subdomain increase the amount of slippage products from a homopolymeric template and, unexpectedly, thumb subdomain deletions decrease the binding affinity for mitochondrial transcription factor (Mtf1). The latter suggests that the thumb subdomain is part of an extended binding surface area involved in binding Mtf1.
authors: a Laboratorio Nacional de Genómica para la Biodiversidad ; Centro de Investigación y de Estudios ; Irapuato , Guanajuato , México.
authors: Gilberto
authors: G
authors: Velazquez
authors: Rui
authors: R
authors: Sousa
authors: Luis G
authors: LG
authors: Brieba
mesh_terms: D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D004278:DNA, Superhelical / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000378:metabolism*; D016368:Frameshift Mutation; D008928:Mitochondria / Q000201:enzymology*; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000378:metabolism*; D061785:Transcription Initiation, Genetic; D014462:Ultrafiltration
chemical_list: D004278:DNA, Superhelical; C071720:MTF1 protein, S cerevisiae; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D012321:DNA-Directed RNA Polymerases; C487996:RPO41 protein, S cerevisiae; D000409:Alanine
keywords: Mtf1, Mitochondrial Transcription Factor; fidelity; mitochondrial RNA polymerase; processivity; ymtRNP, yeast mitochondrial RNA polymerase
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