DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling.

Journal Article

DNA primase synthesizes short RNA primers that initiate DNA synthesis of Okazaki fragments on the lagging strand by DNA polymerase during DNA replication. The binding of prokaryotic DnaG-like primases to DNA occurs at a specific trinucleotide recognition sequence. It is a pivotal step in the formation of Okazaki fragments. Conventional biochemical tools that are used to determine the DNA recognition sequence of DNA primase provide only limited information. Using a high-throughput microarray-based binding assay and consecutive biochemical analyses, it has been shown that 1) the specific binding context (flanking sequences of the recognition site) influences the binding strength of the DNA primase to its template DNA, and 2) stronger binding of primase to the DNA yields longer RNA primers, indicating higher processivity of the enzyme. This method combines PBM and primase activity assay and is designated as high-throughput primase profiling (HTPP), and it allows characterization of specific sequence recognition by DNA primase in unprecedented time and scalability.

Full Text

Duke Authors

Cited Authors

  • Ilic, S; Cohen, S; Afek, A; Gordan, R; Lukatsky, DB; Akabayov, B

Published Date

  • October 8, 2019

Published In

PubMed ID

  • 31657797

Pubmed Central ID

  • 31657797

Electronic International Standard Serial Number (EISSN)

  • 1940-087X

Digital Object Identifier (DOI)

  • 10.3791/59737

Language

  • eng

Conference Location

  • United States