title: DNA unwinding by ring-shaped T4 helicase gp41 is hindered by tension on the occluded strand.
abstract: The replicative helicase for bacteriophage T4 is gp41, which is a ring-shaped hexameric motor protein that achieves unwinding of dsDNA by translocating along one strand of ssDNA while forcing the opposite strand to the outside of the ring. While much study has been dedicated to the mechanism of binding and translocation along the ssDNA strand encircled by ring-shaped helicases, relatively little is known about the nature of the interaction with the opposite, 'occluded' strand. Here, we investigate the interplay between the bacteriophage T4 helicase gp41 and the ss/dsDNA fork by measuring, at the single-molecule level, DNA unwinding events on stretched DNA tethers in multiple geometries. We find that gp41 activity is significantly dependent on the geometry and tension of the occluded strand, suggesting an interaction between gp41 and the occluded strand that stimulates the helicase. However, the geometry dependence of gp41 activity is the opposite of that found previously for the E. coli hexameric helicase DnaB. Namely, tension applied between the occluded strand and dsDNA stem inhibits unwinding activity by gp41, while tension pulling apart the two ssDNA tails does not hinder its activity. This implies a distinct variation in helicase-occluded strand interactions among superfamily IV helicases, and we propose a speculative model for this interaction that is consistent with both the data presented here on gp41 and the data that had been previously reported for DnaB.
authors.affiliation: Department of Physics, University of California Santa Barbara, Santa Barbara, California, United States of America.
authors.forename: Noah
authors.initials: N
authors.lastname: Ribeck
authors.forename: Omar A
authors.initials: OA
authors.lastname: Saleh
mesh_terms: D017122:Bacteriophage T4 / Q000201:enzymology; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics; D053488:DnaB Helicases / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology; D009690:Nucleic Acid Conformation; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*
chemical_list: D004277:DNA, Single-Stranded; D014764:Viral Proteins; C063983:gene 41 protein, Enterobacteria phage T4; D004247:DNA; C506730:dnaB protein, E coli; D004265:DNA Helicases; D053488:DnaB Helicases