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Regulation of DNA replication at the end of the mitochondrial D-loop involves the helicase TWINKLE and a conserved sequence element

Journal article
Authors Elisabeth Jemt
Örjan Persson
Yonghong Shi
Majda Mehmedovic
Jay Uhler
Marcela Davila Lopez
C. Freyer
Claes M Gustafsson
Tore Samuelsson
Maria Falkenberg
Published in Nucleic Acids Research
Volume 43
Issue 19
Pages 9262-9275
ISSN 0305-1048
Publication year 2015
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 9262-9275
Language en
Links dx.doi.org/10.1093/nar/gkv804
Keywords MOUSE L-CELLS, LIGHT-STRAND PROMOTER, TRANSCRIPTION TERMINATION, POLYMERASE-GAMMA, INITIATION SEQUENCE, PRIMER FORMATION, LAGGING-STRAND, HEAVY-STRAND, RNA, REGION, Biochemistry & Molecular Biology, ANG DD, 1985, EMBO JOURNAL, V4, P1559, PPER DP, 1981, JOURNAL OF BIOLOGICAL CHEMISTRY, V256, P5109, ANG DD, 1984, CELL, V36, P635, ARNES MC, 1987, JOURNAL OF BIOLOGICAL CHEMISTRY, V262, P988, ATES OF AMERICA, V107, P16072, ANG DD, 1985, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF
Subject categories Biochemistry and Molecular Biology

Abstract

The majority of mitochondrial DNA replication events are terminated prematurely. The nascent DNA remains stably associated with the template, forming a triple-stranded displacement loop (D-loop) structure. However, the function of the D-loop region of the mitochondrial genome remains poorly understood. Using a comparative genomics approach we here identify two closely related 15 nt sequence motifs of the D-loop, strongly conserved among vertebrates. One motif is at the D-loop 5'-end and is part of the conserved sequence block 1 (CSB1). The other motif, here denoted coreTAS, is at the D-loop 3'-end. Both these sequences may prevent transcription across the D-loop region, since light and heavy strand transcription is terminated at CSB1 and coreTAS, respectively. Interestingly, the replication of the nascent D-loop strand, occurring in a direction opposite to that of heavy strand transcription, is also terminated at coreTAS, suggesting that coreTAS is involved in termination of both transcription and replication. Finally, we demonstrate that the loading of the helicase TWINKLE at coreTAS is reversible, implying that this site is a crucial component of a switch between D-loop formation and full-length mitochondrial DNA replication.

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