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TitleThe human primosome requires replication protein A when copying DNA with inverted repeats.
Journal, issue, pagesNucleic Acids Res, Vol. 53, Issue 15, Year 2025
Publish dateAug 11, 2025
AuthorsAndrey G Baranovskiy / Lucia M Morstadt / Eduardo E Romero / Nigar D Babayeva / Tahir H Tahirov /
PubMed AbstractThe human primosome, a four-subunit complex of primase and DNA polymerase alpha (Polα), initiates DNA synthesis on both chromosome strands by generating chimeric RNA-DNA primers for loading DNA ...The human primosome, a four-subunit complex of primase and DNA polymerase alpha (Polα), initiates DNA synthesis on both chromosome strands by generating chimeric RNA-DNA primers for loading DNA polymerases delta and epsilon (Polϵ). Replication protein A (RPA) tightly binds to single-stranded DNA strands, protecting them from nucleolytic digestion and unauthorized transactions. We report here that RPA plays a critical role for the human primosome during DNA synthesis across inverted repeats prone to hairpin formation. On other alternatively structured DNA, forming a G-quadruplex, RPA does not assist primosome. A stimulatory effect of RPA on DNA synthesis across hairpins was also observed for the catalytic domain of Polα but not of Polϵ. The winged helix-turn-helix domain of RPA is essential for an efficient hairpin bypass and increases RPA-Polα cooperativity on the primed DNA template. Cryo-EM studies revealed that this domain is mainly responsible for the interaction between RPA and Polα. The flexible mode of RPA-Polα interaction during DNA synthesis implies the mechanism of template handover between them when the hairpin formation should be avoided. This work provides insight into a cooperative action of RPA and primosome on DNA, which is critical for DNA synthesis across inverted repeats.
External linksNucleic Acids Res / PubMed:40829805 / PubMed Central
MethodsEM (single particle)
Resolution3.5 Å
Structure data

48312

9mj5

EMDB-48312, PDB-9mj5:
Catalytic domain of human DNA polymerase alpha in complex with DNA and RPA
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

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

Source
  • homo sapiens (human)
KeywordsReplication/DNA/RNA / DNA replication / Replication-DNA-RNA complex

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