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Mammalian transcription factor A is a core component of the mitochondrial transcription machinery

Yonghong Shi ; Anke Dierckx (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; Paulina Wanrooij ; Sjoerd Wanrooij ; N. G. Larsson ; Marcus Wilhelmsson (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; Maria Falkenberg ; Claes M Gustafsson
Proceedings of the National Academy of Science of the United States of America (0027-8424). Vol. 109 (2012), 41, p. 16510-16515.
[Artikel, refereegranskad vetenskaplig]

Transcription factor A (TFAM) functions as a DNA packaging factor in mammalian mitochondria. TFAM also binds sequence-specifically to sites immediately upstream of mitochondrial promoters, but there are conflicting data regarding its role as a core component of the mitochondrial transcription machinery. We here demonstrate that TFAM is required for transcription in mitochondrial extracts as well as in a reconstituted in vitro transcription system. The absolute requirement of TFAM can be relaxed by conditions that allow DNA breathing, i.e., low salt concentrations or negatively supercoiled DNA templates. The situation is thus very similar to that described in nuclear RNA polymerase II-dependent transcription, in which the free energy of supercoiling can circumvent the need for a subset of basal transcription factors at specific promoters. In agreement with these observations, we demonstrate that TFAM has the capacity to induce negative supercoils in DNA, and, using the recently developed nucleobase analog FRET-pair tC(O)-tC(nitro), we find that TFAM distorts significantly the DNA structure. Our findings differ from recent observations reporting that TFAM is not a core component of the mitochondrial transcription machinery. Instead, our findings support a model in which TFAM is absolutely required to recruit the transcription machinery during initiation of transcription.



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Denna post skapades 2012-12-07. Senast ändrad 2015-08-25.
CPL Pubid: 167307

 

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Institutioner (Chalmers)

Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi (GU)
Institutionen för kemi- och bioteknik, Fysikalisk kemi (2005-2014)

Ämnesområden

Livsvetenskaper
Nanovetenskap och nanoteknik
Biofysikalisk kemi
Spektroskopi
Biokemi
Molekylärbiologi

Chalmers infrastruktur

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Denna publikation ingår i:


Fluorescent Nucleobase Analogues and their use for Investigating DNA Interactions


 


Projekt

Denna publikation är ett resultat av följande projekt:


The enzymatic machinery of human mitochondrial dna maintenance (REPMIT) (EC/FP7/261248)