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Human mitochondrial transcription revisited: only TFAM and TFB2M are required for transcription of the mitochondrial genes in vitro.

Journal article
Authors Dmitry Litonin
Marina Sologub
Yonghong Shi
Maria Savkina
Michael Anikin
Maria Falkenberg
Claes M Gustafsson
Dmitry Temiakov
Published in The Journal of biological chemistry
ISSN 1083-351X
Publication year 2010
Published at Institute of Biomedicine
Language en
Links dx.doi.org/10.1074/jbc.C110.128918
Keywords DNA-protein interaction, Mitochondria, RNA polymerase, Transcription, Transcription promoter
Subject categories Chemistry

Abstract

Human mitochondrial transcription is driven by a single subunit RNA polymerase and a set of basal transcription factors. The development of a recombinant in vitro transcription system has allowed for a detailed molecular characterization of the individual components and their contribution to transcription initiation. We found that TFAM and TFB2M act synergistically and increase transcription efficiency 100- to 200-fold compared with RNAP alone. Both the LSP and HSP1 promoters displayed maximal levels of in vitro transcription when TFAM was present in an amount equimolar to the DNA template. Importantly, we did not detect any significant transcription activity in the presence of the TFB2M paralogue, TFB1M or when templates containing the putative HSP2 promoter were used. These data confirm previous observations that TFB1M does not function as a bona fide transcription factor and raise questions as to whether HSP2 serves as a functional promoter in vivo. In addition, we did not detect transcription stimulation by the ribosomal protein MRPL12. Thus, only two essential initiation factors, TFAM and TFB2M, and two promoters, LSP and HSP1, are required to drive transcription of the mitochondrial genome.

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