MGME1 processes flaps into ligatable nicks in concert with DNA polymerase gamma during mtDNA replication
Journal article, 2016

Recently, MGME1 was identified as a mitochondrial DNA nuclease with preference for single-stranded DNA (ssDNA) substrates. Loss-of-function mutations in patients lead to mitochondrial disease with DNA depletion, deletions, duplications and rearrangements. Here, we assess the biochemical role of MGME1 in the processing of flap intermediates during mitochondrial DNA replication using reconstituted systems. We show that MGME1 can cleave flaps to enable efficient ligation of newly replicated DNA strands in combination with POL gamma. MGME1 generates a pool of imprecisely cut products (short flaps, nicks and gaps) that are converted to ligatable nicks by POL gamma through extension or excision of the 3'-end strand. This is dependent on the 3'-5' exonuclease activity of POL gamma which limits strand displacement activity and enables POL gamma to back up to the nick by 3'-5' degradation. We also demonstrate that POL gamma-driven strand displacement is sufficient to generate DNA- but not RNA-flap substrates suitable for MGME1 cleavage and ligation during replication. Our findings have implications for RNA primer removal models, the 5'-end processing of nascent DNA at OriH, and DNA repair.

exonuclease

disease

transfer-rna synthetases

ligase

mice

excision-repair

iii

Biochemistry & Molecular Biology

accessory subunit

endonuclease

okazaki fragment maturation

human mitochondrial-dna

base

Author

Jay Uhler

University of Gothenburg

Christian Thörn

University of Gothenburg

Thomas J. Nicholls

University of Gothenburg

S. Matic

D. Milenkovic

Claes M Gustafsson

University of Gothenburg

Maria Falkenberg

University of Gothenburg

Nucleic Acids Research

0305-1048 (ISSN) 1362-4962 (eISSN)

Vol. 44 12 5861-5871

Subject Categories

Clinical Medicine

DOI

10.1093/nar/gkw468

More information

Created

10/10/2017