Biochemical analysis of human POLG2 variants associated with mitochondrial disease
Open accessAbstract
Defects in mitochondrial DNA (mtDNA) maintenance comprise an expanding repertoire of polymorphic diseases caused, in part, by mutations in the genes encoding the p140 mtDNA polymerase (POLG), its p55 accessory subunit (POLG2) or the mtDNA helicase (C10orf2). In an exploration of nuclear genes for mtDNA maintenance linked to mitochondrial disease, eight heterozygous mutations (six novel) in POLG2 were identified in one control and eight patients with POLG-related mitochondrial disease that lacked POLG mutations. Of these eight mutations, we biochemically characterized seven variants [c.307G>A (G103S); c.457C>G (L153V); c.614C>G (P205R); c.1105A>G (R369G); c.1158T>G (D386E); c.1268C>A (S423Y); c.1423_1424delTT (L475DfsX2)] that were previously uncharacterized along with the wild-type protein and the G451E pathogenic variant. These seven mutations encode amino acid substitutions that map throughout the protein, including the p55 dimer interface and the C-terminal domain that interacts with the catalytic subunit. Recombinant proteins harboring these alterations were assessed for stimulation of processive DNA synthesis, binding to the p140 catalytic subunit, binding to dsDNA and self-dimerization. Whereas the G103S, L153V, D386E and S423Y proteins displayed wild-type behavior, the P205R and R369G p55 variants had reduced stimulation of processivity and decreased affinity for the catalytic subunit. Additionally, the L475DfsX2 variant, which possesses a C-terminal truncation, was unable to bind the p140 catalytic subunit, unable to bind dsDNA and formed aberrant oligomeric complexes. Our biochemical analysis helps explain the pathogenesis of POLG2 mutations in mitochondrial disease and emphasizes the need to quantitatively characterize the biochemical consequences of newly discovered mutations before classifying them as pathogenic.
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Of the handful of heterozygous mutations identified in POLG2, G451E and R369G are linked to PEO.
“Of the handful of heterozygous mutations identified in POLG2, G451E (Longley et al., 2006) and R369G (Craig et al., 2012; Young et al., 2011) are linked to PEO.”
From Role of the mitochondrial DNA replication machinery in mitochondrial DNA mutagenesis, aging and age-related diseases (2017) · cites this paper as Young et al., 2011
G451E homodimers fail to enhance binding of PolG to DNA, while R369G has weaker binding to PolG.
“G451E homodimers also fail to enhance binding of PolG to DNA, while R369G has weaker binding to PolG.”
From Role of the mitochondrial DNA replication machinery in mitochondrial DNA mutagenesis, aging and age-related diseases (2017) · cites this paper as Young et al., 2011
Homodimeric mutants have problems stimulating PolG processivity, with G451E being more severely defective than R396G.
“Homodimeric mutants have problems stimulating PolG processivity, with G451E being more severely defective than R396G (Craig et al., 2012; Longley et al., 2006; Young et al., 2011).”
From Role of the mitochondrial DNA replication machinery in mitochondrial DNA mutagenesis, aging and age-related diseases (2017) · cites this paper as Young et al., 2011
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- Authors
- Young, M. J., Longley, M. J., Li, F.-Y., Kasiviswanathan, R., Wong, L.-J., & Copeland, W. C.
- Affiliations
- National Institutes of Health (US)
- Baylor College of Medicine (US)
- Corresponding author
- Wong, Lee-Jun
- Copeland, William C.
- Venue
- Human Molecular Genetics
- Publisher
- Oxford University Press (OUP)
- Published
- 2011-05-09
- Volume
- 20
- Issue
- 15
- Pages
- 3052-3066
- ISSN
- 0964-6906 (ISSN-L)
- 1460-2083
- PMC
- PMC3131046
- OpenAlex
- W2107759081
- Cited by
- 66
- References
- 46
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 3 · 2023: 4 · 2022: 1 · 2021: 1 · 2020: 6 · 2019: 3 · 2018: 5 · 2017: 4 · 2016: 9 · 2015: 9 · 2014: 7 · 2013: 3 · 2012: 3
- FWCI
- 2.21
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Human Molecular Genetics (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Environmental Health Sciences (ES-065078)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genomics and Rare Diseases
- Keywords
- POLG2, mitochondrial DNA maintenance, mitochondrial disease, novel mutations, amino acid substitutions, processive DNA synthesis, protein dimerization, C-terminal truncation, pathogenicity classification
- MeSH terms
- Chromatography, Gel, DNA-Directed DNA Polymerase, Humans, Molecular Sequence Data, Mutation, Blotting, Western, Protein Structure, Secondary, Amino Acid Substitution, Mitochondrial Diseases, Immunoprecipitation
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