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TitleAn interaction network in the polymerase active site is a prerequisite for Watson-Crick base pairing in Pol γ.
Journal, issue, pagesSci Adv, Vol. 10, Issue 21, Page eadl3214, Year 2024
Publish dateMay 24, 2024
AuthorsJoon Park / Geoffrey K Herrmann / Arkanil Roy / Christie K Shumate / G Andrés Cisneros / Y Whitney Yin /
PubMed AbstractThe replication accuracy of DNA polymerase gamma (Pol γ) is essential for mitochondrial genome integrity. Mutation of human Pol γ arginine-853 has been linked to neurological diseases. Although not ...The replication accuracy of DNA polymerase gamma (Pol γ) is essential for mitochondrial genome integrity. Mutation of human Pol γ arginine-853 has been linked to neurological diseases. Although not a catalytic residue, Pol γ arginine-853 mutants are void of polymerase activity. To identify the structural basis for the disease, we determined a crystal structure of the Pol γ mutant ternary complex with correct incoming nucleotide 2'-deoxycytidine 5'-triphosphate (dCTP). Opposite to the wild type that undergoes open-to-closed conformational changes when bound to a correct nucleotide that is essential for forming a catalytically competent active site, the mutant complex failed to undergo the conformational change, and the dCTP did not base pair with its Watson-Crick complementary templating residue. Our studies revealed that arginine-853 coordinates an interaction network that aligns the 3'-end of primer and dCTP with the catalytic residues. Disruption of the network precludes the formation of Watson-Crick base pairing and closing of the active site, resulting in an inactive polymerase.
External linksSci Adv / PubMed:38787958 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.37 - 3.43 Å
Structure data

42150

8udl

EMDB-42150, PDB-8udl:
Human Mitochondrial DNA Polymerase Gamma Binary Complex
Method: EM (single particle) / Resolution: 2.37 Å

PDB-8udk:
Human Mitochondrial DNA Polymerase gamma R853A Ternary Complex
Method: X-RAY DIFFRACTION / Resolution: 3.43 Å

Chemicals

ChemComp-DCP:
2'-DEOXYCYTIDINE-5'-TRIPHOSPHATE

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsTRANSFERASE/DNA / Mitochondrial / DNA Polymerase / TRANSFERASE / TRANSFERASE-DNA complex

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