title: Herpes simplex-1 helicase-primase. Identification of two nucleoside triphosphatase sites that promote DNA helicase action.
abstract: Herpes simplex virus type 1 (HSV-1) encodes a heterotrimeric helicase-primase comprised of the products of the UL5, UL8, and UL52 genes (Crute, J. J., and Lehman, I. R. (1991) J. Biol. Chem. 266, 4484-4488). A steady state kinetic analysis of the enzyme isolated from HSV-1-infected CV-1 cells or insect cells expressing the enzyme after infection with recombinant baculoviruses has shown it to possess two sites capable of hydrolyzing nucleoside triphosphates in a DNA-dependent manner. One site (Site I) hydrolyzes both ATP and GTP; the second (Site II) hydrolyzes only ATP. These two sites are contained within a subassembly of the helicase-primase formed by coexpression of the UL5 and UL52 genes in insect cells. Sites I and II are activated by separate DNA effector sites, both of which support DNA helicase action. These findings are likely to be of importance in understanding how helicases in general catalyze the unwinding of duplex DNA and, in particular, how the helicase-primase functions at the HSV-1 replication fork.
authors.affiliation: Department of Biochemistry, Beckman Center, Stanford University School of Medicine, California 94305-5307.
authors.forename: J J
authors.initials: JJ
authors.lastname: Crute
authors.forename: R C
authors.initials: RC
authors.lastname: Bruckner
authors.forename: M S
authors.initials: MS
authors.lastname: Dodson
authors.forename: I R
authors.initials: IR
authors.lastname: Lehman
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D019915:DNA Primase; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D018139:Simplexvirus / Q000201:enzymology*; D014764:Viral Proteins
chemical_list: D046911:Macromolecular Substances; D014764:Viral Proteins; D004247:DNA; D019915:DNA Primase; C062021:helicase-primase, Human herpesvirus 1; D000251:Adenosine Triphosphatases; D020558:GTP Phosphohydrolases; D004265:DNA Helicases