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TitleStructures of the mitochondrial single-stranded DNA binding protein with DNA and DNA polymerase γ.
Journal, issue, pagesNucleic Acids Res, Vol. 52, Issue 17, Page 10329-10340, Year 2024
Publish dateSep 23, 2024
AuthorsAmanda A Riccio / Jonathan Bouvette / Lars C Pedersen / Shruti Somai / Robert C Dutcher / Mario J Borgnia / William C Copeland /
PubMed AbstractThe mitochondrial single-stranded DNA (ssDNA) binding protein, mtSSB or SSBP1, binds to ssDNA to prevent secondary structures of DNA that could impede downstream replication or repair processes. ...The mitochondrial single-stranded DNA (ssDNA) binding protein, mtSSB or SSBP1, binds to ssDNA to prevent secondary structures of DNA that could impede downstream replication or repair processes. Clinical mutations in the SSBP1 gene have been linked to a range of mitochondrial disorders affecting nearly all organs and systems. Yet, the molecular determinants governing the interaction between mtSSB and ssDNA have remained elusive. Similarly, the structural interaction between mtSSB and other replisome components, such as the mitochondrial DNA polymerase, Polγ, has been minimally explored. Here, we determined a 1.9-Å X-ray crystallography structure of the human mtSSB bound to ssDNA. This structure uncovered two distinct DNA binding sites, a low-affinity site and a high-affinity site, confirmed through site-directed mutagenesis. The high-affinity binding site encompasses a clinically relevant residue, R38, and a highly conserved DNA base stacking residue, W84. Employing cryo-electron microscopy, we confirmed the tetrameric assembly in solution and capture its interaction with Polγ. Finally, we derived a model depicting modes of ssDNA wrapping around mtSSB and a region within Polγ that mtSSB binds.
External linksNucleic Acids Res / PubMed:39106165 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.9 - 3.5 Å
Structure data

EMDB-42842: A Mitochondrial Replication Complex: PolG/PolG2 Bound to DNA in complex with the Single-Stranded Binding Protein (mtSSB)
Method: EM (single particle) / Resolution: 3.5 Å

PDB-8uzt:
Mitochondrial single-stranded binding protein bound to DNA
Method: X-RAY DIFFRACTION / Resolution: 1.9 Å

Chemicals

ChemComp-GOL:
GLYCEROL

ChemComp-NA:
Unknown entry

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN/DNA / single-stranded binding protein / DNA BINDING PROTEIN / mitochondrial SSB / SSB / mitochondrial replication protein / DNA BINDING PROTEIN-DNA complex

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