Characterization of a 5'-polynucleotide kinase/3'-phosphatase from bacteriophage RM378.
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.
Blondal
Thorarinn
T
Prokaria Limited. Gylfaflot 5, 112 Reyjavik, Iceland.
Hjorleifsdottir
Sigridur
S
Aevarsson
Arnthor
A
Fridjonsson
Olafur H
OH
Skirnisdottir
Sigurlaug
S
Wheat
Jon Oskar
JO
Hermannsdottir
Anna Gudny
AG
Hreggvidsson
Gudmundur O
GO
Smith
Albert Vernon
AV
Kristjansson
Jakob K
JK
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
D000244:Adenosine Diphosphate; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D010744:Phosphoric Monoester Hydrolases