title: Energy-independent helicase activity of a viral genome packaging motor.
abstract: The assembly of complex double-stranded DNA viruses includes a genome packaging step where viral DNA is translocated into the confines of a preformed procapsid shell. In most cases, the preferred packaging substrate is a linear concatemer of viral genomes linked head-to-tail. Viral terminase enzymes are responsible for both excision of an individual genome from the concatemer (DNA maturation) and translocation of the duplex into the capsid (DNA packaging). Bacteriophage λ terminase site-specifically nicks viral DNA at the cos site in a concatemer and then physically separates the nicked, annealed strands to mature the genome in preparation for packaging. Here we present biochemical studies on the so-called helicase activity of λ terminase. Previous studies reported that ATP is required for strand separation, and it has been presumed that ATP hydrolysis is required to drive the reaction. We show that ADP and nonhydrolyzable ATP analogues also support strand separation at low (micromolar) concentrations. In addition, the Escherichia coli integration host factor protein (IHF) strongly stimulates the reaction in a nucleotide-independent manner. Finally, we show that elevated concentrations of nucleotide inhibit both ATP- and IHF-stimulated strand separation by λ terminase. We present a model where nucleotide and IHF interact with the large terminase subunit and viral DNA, respectively, to engender a site-specifically bound, catalytically competent genome maturation complex. In contrast, binding of nucleotide to the low-affinity ATP binding site in the small terminase subunit mediates a conformational switch that down-regulates maturation activities and activates the DNA packaging activity of the enzyme. This affords a motor complex that binds tightly, but nonspecifically, to DNA as it translocates the duplex into the capsid shell. These studies have yielded mechanistic insight into the assembly of the maturation complex on viral DNA and its transition to a mobile packaging motor that may be common to all of the complex double-stranded DNA viruses.
authors.affiliation: Department of Medicinal Chemistry, School of Pharmacy, University of Washington, Seattle, Washington 98195-7610, United States.
authors.forename: Jenny R
authors.initials: JR
authors.lastname: Chang
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Andrews
authors.forename: Carlos E
authors.initials: CE
authors.lastname: Catalano
mesh_terms: D000256:Adenoviridae / Q000201:enzymology / Q000235:genetics; D017103:Bacillus Phages / Q000201:enzymology / Q000235:genetics; D010582:Bacteriophage lambda / Q000201:enzymology* / Q000235:genetics; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics; D004734:Energy Metabolism / Q000235:genetics; D016679:Genome, Viral; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000737:chemistry* / Q000235:genetics; D019065:Virus Assembly / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D020409:Molecular Motor Proteins; D004265:DNA Helicases