title: Characterization of a 5'-polynucleotide kinase/3'-phosphatase from bacteriophage RM378.
abstract: A polynucleotide kinase from the thermophilic bacteriophage RM378 that infects the thermophilic eubacterium Rhodothermus marinus was identified, expressed, and purified. This polynucleotide kinase was demonstrated to have a 5'-kinase domain as well as a 3'-phosphohydrolase domain. The RM378 polynucleotide kinase had limited sequence similarity to the 5'-kinase domain of the T4 bacteriophage polynucleotide kinase, but apparent homology was not evident within the 3'-phosphohydrolase domain. The domain order of RM378 polynucleotide kinase was reversed relative to that of the T4 polynucleotide kinase. The RM378 phosphohydrolase domain displayed some sequence similarity with the bacterial poly(A) polymerase family, including an HD motif characteristic of the diverse superfamily of metal-dependent HD phosphohydrolases. The RM378 polynucleotide kinase was biochemically characterized and shown to possess 5'-kinase activity on RNA and single- and double-stranded DNA at elevated temperatures. It also showed phosphohydrolase activity on 2':3'-cyclic adenosine monophosphate. This description of the RM378 polynucleotide kinase, along with the recently described RM378 RNA ligase, suggests that the RM378 bacteriophage has to counter a similar anti-phage mechanism in R. marinus as the one that the T4 phage has to counter in Escherichia coli.
authors.affiliation: Prokaria Limited. Gylfaflot 5, 112 Reyjavik, Iceland.
authors.forename: Thorarinn
authors.initials: T
authors.lastname: Blondal
authors.forename: Sigridur
authors.initials: S
authors.lastname: Hjorleifsdottir
authors.forename: Arnthor
authors.initials: A
authors.lastname: Aevarsson
authors.forename: Olafur H
authors.initials: OH
authors.lastname: Fridjonsson
authors.forename: Sigurlaug
authors.initials: S
authors.lastname: Skirnisdottir
authors.forename: Jon Oskar
authors.initials: JO
authors.lastname: Wheat
authors.forename: Anna Gudny
authors.initials: AG
authors.lastname: Hermannsdottir
authors.forename: Gudmundur O
authors.initials: GO
authors.lastname: Hreggvidsson
authors.forename: Albert Vernon
authors.initials: AV
authors.lastname: Smith
authors.forename: Jakob K
authors.initials: JK
authors.lastname: Kristjansson
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
chemical_list: D000244:Adenosine Diphosphate; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D010744:Phosphoric Monoester Hydrolases