Publication Date:
2011-03-11
Description:
Mammalian cells have three ATP-dependent DNA ligases, which are required for DNA replication and repair. Homologues of ligase I (Lig1) and ligase IV (Lig4) are ubiquitous in Eukarya, whereas ligase III (Lig3), which has nuclear and mitochondrial forms, appears to be restricted to vertebrates. Lig3 is implicated in various DNA repair pathways with its partner protein Xrcc1 (ref. 1). Deletion of Lig3 results in early embryonic lethality in mice, as well as apparent cellular lethality, which has precluded definitive characterization of Lig3 function. Here we used pre-emptive complementation to determine the viability requirement for Lig3 in mammalian cells and its requirement in DNA repair. Various forms of Lig3 were introduced stably into mouse embryonic stem (mES) cells containing a conditional allele of Lig3 that could be deleted with Cre recombinase. With this approach, we find that the mitochondrial, but not nuclear, Lig3 is required for cellular viability. Although the catalytic function of Lig3 is required, the zinc finger (ZnF) and BRCA1 carboxy (C)-terminal-related (BRCT) domains of Lig3 are not. Remarkably, the viability requirement for Lig3 can be circumvented by targeting Lig1 to the mitochondria or expressing Chlorella virus DNA ligase, the minimal eukaryal nick-sealing enzyme, or Escherichia coli LigA, an NAD(+)-dependent ligase. Lig3-null cells are not sensitive to several DNA-damaging agents that sensitize Xrcc1-deficient cells. Our results establish a role for Lig3 in mitochondria, but distinguish it from its interacting protein Xrcc1.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3261757/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3261757/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Simsek, Deniz -- Furda, Amy -- Gao, Yankun -- Artus, Jerome -- Brunet, Erika -- Hadjantonakis, Anna-Katerina -- Van Houten, Bennett -- Shuman, Stewart -- McKinnon, Peter J -- Jasin, Maria -- CA21765/CA/NCI NIH HHS/ -- ES019566/ES/NIEHS NIH HHS/ -- GM54668/GM/NIGMS NIH HHS/ -- NS37956/NS/NINDS NIH HHS/ -- P30 CA008748/CA/NCI NIH HHS/ -- R01 ES019566/ES/NIEHS NIH HHS/ -- R01 GM054668/GM/NIGMS NIH HHS/ -- R01 GM054668-12/GM/NIGMS NIH HHS/ -- R01 GM054668-12S1/GM/NIGMS NIH HHS/ -- R01 GM063611/GM/NIGMS NIH HHS/ -- R01 NS037956/NS/NINDS NIH HHS/ -- R01 NS037956-13/NS/NINDS NIH HHS/ -- England -- Nature. 2011 Mar 10;471(7337):245-8. doi: 10.1038/nature09794.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21390132" target="_blank"〉PubMed〈/a〉
Keywords:
Animals
;
Biocatalysis
;
Cell Survival
;
DNA Damage
;
DNA Ligases/chemistry/deficiency/genetics/*metabolism
;
*DNA Repair
;
DNA, Mitochondrial/*metabolism
;
DNA-Binding Proteins/*metabolism
;
Embryonic Stem Cells/metabolism
;
Genes, Essential
;
Genetic Complementation Test
;
Humans
;
Mice
;
Mitochondria/*enzymology/*genetics/pathology
;
Protein Structure, Tertiary
;
Sister Chromatid Exchange/drug effects
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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