Skip to main content

Functional Coupling of Duplex Translocation to DNA Cleavage in a Type I Restriction Enzyme

Author(s): Csefalvay, Eva; Lapkouski, Mikalai; Guzanova, Alena; Csefalvay, Ladislav; Baikova, Tatsiana; et al

To refer to this page use:
Abstract: Type I restriction-modification enzymes are multifunctional heteromeric complexes with DNA cleavage and ATP-dependent DNA translocation activities located on motor subunit HsdR. Functional coupling of DNA cleavage and translocation is a hallmark of the Type I restriction systems that is consistent with their proposed role in horizontal gene transfer. DNA cleavage occurs at nonspecific sites distant from the cognate recognition sequence, apparently triggered by stalled translocation. The X-ray crystal structure of the complete HsdR subunit from E. coli plasmid R124 suggested that the triggering mechanism involves interdomain contacts mediated by ATP. In the present work, in vivo and in vitro activity assays and crystal structures of three mutants of EcoR124I HsdR designed to probe this mechanism are reported. The results indicate that interdomain engagement via ATP is indeed responsible for signal transmission between the endonuclease and helicase domains of the motor subunit. A previously identified sequence motif that is shared by the RecB nucleases and some Type I endonucleases is implicated in signaling. © 2015 Csefalvay et al.
Publication Date: 3-Jun-2015
Electronic Publication Date: 3-Jun-2015
Citation: Csefalvay, Eva, Lapkouski, Mikalai, Guzanova, Alena, Csefalvay, Ladislav, Baikova, Tatsiana, Shevelev, Igor, Bialevich, Vitali, Shamayeva, Katsiaryna, Janscak, Pavel, Kuta Smatanova, Ivana, Panjikar, Santosh, Carey, Jannette, Weiserova, Marie, Ettrich, Rüdiger. (2015). Functional Coupling of Duplex Translocation to DNA Cleavage in a Type I Restriction Enzyme. PLOS ONE, 10 (6), e0128700 - e0128700. doi:10.1371/journal.pone.0128700.
DOI: doi:10.1371/journal.pone.0128700
EISSN: 1932-6203
Pages: 10.6:e0128700-1 - e0128700-21
Type of Material: Journal Article
Journal/Proceeding Title: PLOS ONE
Version: Final published version. This is an open access article.
Notes: PLoS ONE. Volume 10, Issue 6, 3 June 2015, Article number e0128700.

Items in OAR@Princeton are protected by copyright, with all rights reserved, unless otherwise indicated.