umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
A two-nuclease pathway involving RNase H1 is required for primer removal at human mitochondrial OriL
Show others and affiliations
2018 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 46, no 18, p. 9471-9483Article in journal (Refereed) Published
Abstract [en]

The role of Ribonuclease H1 (RNase H1) during primer removal and ligation at the mitochondrial origin of light-strand DNA synthesis (OriL) is a key, yet poorly understood, step in mitochondrial DNA maintenance. Here, we reconstitute the replication cycle of L-strand synthesis in vitro using recombinant mitochondrial proteins and model OriL substrates. The process begins with initiation of DNA replication at OriL and ends with primer removal and ligation. We find that RNase H1 partially removes the primer, leaving behind the last one to three ribonucleotides. These 5′-end ribonucleotides disturb ligation, a conclusion which is supported by analysis of RNase H1-deficient patient cells. A second nuclease is therefore required to remove the last ribonucleotides and we demonstrate that Flap endonuclease 1 (FEN1) can execute this function in vitro. Removal of RNA primers at OriL thus depends on a two-nuclease model, which in addition to RNase H1 requires FEN1 or a FEN1-like activity. These findings define the role of RNase H1 at OriL and help to explain the pathogenic consequences of disease causing mutations in RNase H1.

Place, publisher, year, edition, pages
Oxford University Press, 2018. Vol. 46, no 18, p. 9471-9483
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-154079DOI: 10.1093/nar/gky708ISI: 000450953200023PubMedID: 30102370OAI: oai:DiVA.org:umu-154079DiVA, id: diva2:1270199
Funder
Swedish Research Council, VR521-2013-3621Swedish Cancer Society, CAN 2016/816Knut and Alice Wallenberg FoundationAvailable from: 2018-12-12 Created: 2018-12-12 Last updated: 2018-12-12Bibliographically approved

Open Access in DiVA

fulltext(9037 kB)45 downloads
File information
File name FULLTEXT01.pdfFile size 9037 kBChecksum SHA-512
f436f457c34ca06fcbfae5e59f947ee1315e8e343d9871653ba3ffa71bc1cde7a580c347f4e62c601ee083597279e10ab38507122458f71f05b155434bd645fc
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Authority records BETA

Doimo, MaraWanrooij, Sjoerd

Search in DiVA

By author/editor
Al-Behadili, AliDoimo, MaraReyes, AurelioWanrooij, SjoerdFalkenberg, Maria
By organisation
Department of Medical Biochemistry and Biophysics
In the same journal
Nucleic Acids Research
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 45 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 535 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf