Defining a novel domain that provides an essential contribution to site-specific interaction of Rep protein with DNA.

ABSTRACT: An essential feature of replication initiation proteins is their ability to bind to DNA. In this work, we describe a new domain that contributes to a replication initiator sequence-specific interaction with DNA. Applying biochemical assays and structure prediction methods coupled with DNA-protein crosslinking, mass spectrometry, and construction and analysis of mutant proteins, we identified that the replication initiator of the broad host range plasmid RK2, in addition to two winged helix domains, contains a third DNA-binding domain. The phylogenetic analysis revealed that the composition of this unique domain is typical within the described TrfA-like protein family. Both in vitro and in vivo experiments involving the constructed TrfA mutant proteins showed that the newly identified domai

SUBMITTER: Wegrzyn K 

PROVIDER: S-EPMC8034659 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature