title: The enzymology of a viral genome packaging motor is influenced by the assembly state of the motor subunits.
abstract: Terminase enzymes are responsible for the excision of a single genome from a concatemeric precursor (genome maturation) and concomitant packaging of DNA into the capsid shell. Here, we demonstrate that lambda terminase can be purified as a homogeneous "protomer" species, and we present a kinetic analysis of the genome maturation and packaging activities of the protomeric enzyme. The protomer assembles into a distinct maturation complex at the cos sequence of a concatemer. This complex rapidly nicks the duplex to form the mature left end of the viral genome, which is followed by procapsid binding, activation of the packaging ATPase, and translocation of the duplex into the capsid interior by the terminase motor complex. Genome packaging by the protomer shows high fidelity with only the mature left end of the duplex inserted into the capsid shell. In sum, the data show that the terminase protomer exhibits catalytic activity commensurate with that expected of a bone fide genome maturation and packaging complex in vivo and that both catalytically competent complexes are composed of four terminase protomers assembled into a ringlike structure that encircles duplex DNA. This work provides mechanistic insight into the coordinated catalytic activities of terminase enzymes in virus assembly that can be generalized to all of the double-stranded DNA viruses.
authors.affiliation: Department of Medicinal Chemistry, School of Pharmacy, University of Washington, Seattle, Washington 98195-7610, United States.
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Andrews
authors.forename: Carlos Enrique
authors.initials: CE
authors.lastname: Catalano
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D042003:DNA Packaging; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D016679:Genome, Viral; D006868:Hydrolysis; D007700:Kinetics; D011401:Promoter Regions, Genetic
chemical_list: D000255:Adenosine Triphosphate; D004706:Endodeoxyribonucleases; C030894:terminase