TY - JOUR
T1 - Yeast mitochondrial RNAP conformational changes are regulated by interactions with the mitochondrial transcription factor
AU - Drakulic, Srdja
AU - Wang, Liping
AU - Cuéllar, Jorge
AU - Guo, Qing
AU - Velázquez, Gilberto
AU - Martín-Benito, Jaime
AU - Sousa, Rui
AU - Valpuesta, José M.
N1 - Funding Information:
Spanish Ministry of Science and Innovation [BFU2010-15703, BFU2013-44202 to J.M.V.]; National Institutes of Health [GM0502522 to R.S.]. La Caixa foundation fellowship [to S.D.].Funding for open access charge: Spanish Ministry of Science and Innovation [BFU2013-44202] [ to J.M.V.]. Conflict of interest statement. None declared.
PY - 2014/6/27
Y1 - 2014/6/27
N2 - Mitochondrial RNA polymerases (MtRNAPs) are members of the single-subunit RNAP family, the most well-characterized member being the RNAP from T7 bacteriophage. MtRNAPs are, however, functionally distinct in that they depend on one or more transcription factors to recognize and open the promoter and initiate transcription, while the phage RNAPs are capable of performing these tasks alone. Since the transcriptional mechanisms that are conserved in phage and mitochondrial RNAPs have been so effectively characterized in the phage enzymes, outstanding structure-mechanism questions concern those aspects that are distinct in the MtRNAPs, particularly the role of the mitochondrial transcription factor(s). To address these questions we have used both negative staining and cryo-EM to generate threedimensional reconstructions of yeast MtRNAP initiation complexes with and without the mitochondrial transcription factor (MTF1), and of the elongation complex. Together with biochemical experiments, these data indicate that MTF1 uses multiple mechanisms to drive promoter opening, and that its interactions with the MtRNAP regulate the conformational changes undergone by the latter enzyme as it traverses the template strand.
AB - Mitochondrial RNA polymerases (MtRNAPs) are members of the single-subunit RNAP family, the most well-characterized member being the RNAP from T7 bacteriophage. MtRNAPs are, however, functionally distinct in that they depend on one or more transcription factors to recognize and open the promoter and initiate transcription, while the phage RNAPs are capable of performing these tasks alone. Since the transcriptional mechanisms that are conserved in phage and mitochondrial RNAPs have been so effectively characterized in the phage enzymes, outstanding structure-mechanism questions concern those aspects that are distinct in the MtRNAPs, particularly the role of the mitochondrial transcription factor(s). To address these questions we have used both negative staining and cryo-EM to generate threedimensional reconstructions of yeast MtRNAP initiation complexes with and without the mitochondrial transcription factor (MTF1), and of the elongation complex. Together with biochemical experiments, these data indicate that MTF1 uses multiple mechanisms to drive promoter opening, and that its interactions with the MtRNAP regulate the conformational changes undergone by the latter enzyme as it traverses the template strand.
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U2 - 10.1093/nar/gku795
DO - 10.1093/nar/gku795
M3 - Article
C2 - 25183523
AN - SCOPUS:84954278133
VL - 42
SP - 11246
EP - 11260
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 17
ER -