Flash DNA sequence context controls the binding and processivity of T7 DNA primase - ICS84 The 84th Annual Meeting of the Israel Chemical Society

Stefan Ilic ¹ Arial Afek ² David Lukatsky ¹ Raluca Gordan ² Barak Akabayov ¹

¹Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel
²Department of Biostatistics and Bioinformatics, Duke University, Durham, NC, USA

Primases are essential enzymes for DNA replication process. They synthesize short RNA primers on a lagging DNA strand that are used by DNA polymerase to form Okazaki fragments. We investigated the DNA-binding preferences and activity of bacteriophage T7 DNA primase by using a new workflow called high-throughput primas profiling (HTPP). This workflow combined microarray-based DNA-binding assay and biochemical analyses to reveal a complex binding specificity and functional activity of T7 DNA primase on various DNA templates. 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). We believe that applying HTPP to other DNA-processing enzymes will give new insights into the effect of DNA sequence composition on DNA-recognition and activity of these enzymes.