Interactions of DNA with Human DNA Primase Monitored with Photoactivatable Cross-Linking Agents: Implications for the Role of the p58 Subunit

1999/09/01 by Bahram Arezi, Brian W. Kirk, William C. Copeland +1 · 47 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · #Biochemistry #Biology #Chemistry #DNA #DNA Repair Mechanisms #DNA and Nucleic Acid Chemistry #DNA polymerase #DNA replication #Gene #Molecular biology #Polymerase chain reaction #Primase #Primer (cosmetics) #Protein subunit #RNA Interference and Gene Delivery #Reverse transcriptase

paper · doi:10.1021/bi9908991

published in Biochemistry 38(39), 12899-12907 (American Chemical Society)

openalex publication_date 1999/09/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01

Abstract

Regulation of the p49-p58 primase complex during primer synthesis and the interaction of the primase subunits with DNA were examined. After primase synthesizes a primer that DNA polymerase alpha (pol alpha) can readily elongate, further primase activity is negatively regulated. This occurs within both the context of the four-subunit pol alpha-primase complex and in the p49-p58 primase complex, indicating that the newly generated primer-template species need not interact with pol alpha to regulate further primase activity. Photo-cross-linking of single-stranded DNA-primase complexes revealed that whereas the isolated p49 and p58 subunits both reacted with DNA upon photolysis, only the p58 subunit reacted with the DNA when photolysis was performed using the p49-p58 primase complex. After primer synthesis by the complex, p58 was again the only subunit that reacted with the DNA. These results suggest a model for regulation of primer synthesis in which the newly synthesized primer-template species binds to p58 and regulates further primer synthesis. Additionally, the ability of p58 to interact with primer-template species suggests that p58 mediates the transfer of primers from the primase active site to pol alpha.

Cited by

• An iron-sulfur domain of the eukaryotic primase is essential for RNA primer synthesis. [europepmc]
• Mechanism and evolution of DNA primases. [europepmc]
• Herpes simplex virus-1 helicase-primase: roles of each subunit in DNA binding and phosphodiester bond formation. [europepmc]
• The archaeo-eukaryotic GINS proteins and the archaeal primase catalytic subunit PriS share a common domain. [europepmc]
• Shared active site architecture between the large subunit of eukaryotic primase and DNA photolyase. [europepmc]
• Essential role of the iron-sulfur cluster binding domain of the primase regulatory subunit Pri2 in DNA replication initiation. [europepmc]
• Primer synthesis by a eukaryotic-like archaeal primase is independent of its Fe-S cluster. [europepmc]
• Substrate Binding Regulates Redox Signaling in Human DNA Primase. [europepmc]
• Yeast require redox switching in DNA primase. [europepmc]
• Functional and structural similarity of human DNA primase [4Fe4S] cluster domain constructs. [europepmc]
• Modification of the 4Fe-4S Cluster Charge Transport Pathway Alters RNA Synthesis by Yeast DNA Primase. [europepmc]
• Structures of human primosome elongation complexes. [europepmc]
• Models for human telomere C-strand fill-in by CST-Polα-primase. [europepmc]
• Flexibility and Distributive Synthesis Regulate RNA Priming and Handoff in Human DNA Polymerase α-Primase. [europepmc]
• A mechanistic model of primer synthesis from catalytic structures of DNA polymerase α-primase. [europepmc]

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