Initial Primer Synthesis of a DNA Primase Monitored by Real-Time NMR Spectroscopy.

ABSTRACT: Primases are crucial enzymes for DNA replication, as they synthesize a short primer required for initiating DNA replication. We herein present time-resolved nuclear magnetic resonance (NMR) spectroscopy in solution and in the solid state to study the initial dinucleotide formation reaction of archaeal pRN1 primase. Our findings show that the helix-bundle domain (HBD) of pRN1 primase prepares the two substrates and then hands them over to the catalytic domain to initiate the reaction. By using nucleotide triphosphate analogues, the reaction is substantially slowed down, allowing us to study the initial dinucleotide formation in real time. We show that the sedimented protein-DNA complex remains active in the solid-state NMR rotor and that time-resolved ³¹P-detected cross-polarizat

SUBMITTER: Wu P 

PROVIDER: S-EPMC11009956 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature