DNA Sequence Context Controls the Binding and Processivity of the T7 DNA Primase

Published

Journal Article

© 2018 The Author(s) Primases are key enzymes involved in DNA replication. They act on single-stranded DNA and catalyze the synthesis of short RNA primers used by DNA polymerases. Here, we investigate the DNA binding and activity of the bacteriophage T7 primase using a new workflow called high-throughput primase profiling (HTPP). Using a unique combination of high-throughput binding assays and biochemical analyses, HTPP reveals a complex landscape of binding specificity and functional activity for the T7 primase, determined by sequences flanking the primase recognition site. We identified specific features, such as G/T-rich flanks, which increase primase-DNA binding up to 10-fold and, surprisingly, also increase the length of newly formed RNA (up to 3-fold). To our knowledge, variability in primer length has not been reported for this primase. We expect that applying HTPP to additional enzymes will reveal new insights into the effects of DNA sequence composition on the DNA recognition and functional activity of primases. Biochemical Mechanism; Molecular Biology; Molecular Genetics

Full Text

Duke Authors

Cited Authors

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

Published Date

  • April 27, 2018

Published In

Volume / Issue

  • 2 /

Start / End Page

  • 141 - 147

Electronic International Standard Serial Number (EISSN)

  • 2589-0042

Digital Object Identifier (DOI)

  • 10.1016/j.isci.2018.03.019

Citation Source

  • Scopus