Novel deoxynucleoside-phosphorylating enzymes in mycoplasmas: evidence for efficient utilization of deoxynucleosides.
Mycoplasmas are unable to synthesize purine and pyrimidine bases de novo. Therefore, salvage of existing nucleosides and bases is essential for their survival. Four mycoplasma species were studied with regard to their ability to phosphorylate deoxynucleosides. High levels of thymidine kinase (TK), deoxycytidine kinase (dCK), deoxyguanosine kinase (dGK) and deoxyadenosine kinase (dAK) activities were detected in extracts from Mycoplasma pneumoniae, Mycoplasma mycoides subsp. mycoides SC (M. mymySC), Acholeplasma laidlawii (A. laidlawii) and Mycoplasma arginini (M. arginini). Nucleoside phosphotransferase activities were found at high levels in A. laidlawii and low levels in M. arginini. Pyrophosphate-dependent deoxynucleoside kinase activities were detected mainly in A. laidlawii and M. mymySC extracts. Two open reading frames were identified in the M. mymySC genome; one showed 25% sequence identity to human dGK and the other one had about 26% sequence identity to human TK1. The M. mymySC dGK-like enzyme was cloned, expressed in Escherichia coli and affinity-purified. This enzyme phosphorylated dAdo, dGuo and dCyd, and the highest catalytic rate was with dAdo as substrate. Therefore, we suggest that this enzyme should be named deoxyadenosine kinase. The physiological role of mycoplasma dAK and TK may be to support the unusually large dATP and dTTP pools required for replication of mycoplasma genomes.
Wang
L
L
Department of Veterinary Medical Chemistry, The Swedish University of Agricultural Sciences, The Biomedical Centre, SE-751 23 Uppsala, Sweden. liya.wang@vmk.slu.se
Westberg
J
J
Bölske
G
G
Eriksson
S
S
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003839:Deoxyadenosines / Q000378:metabolism; D003841:Deoxycytidine / Q000378:metabolism; D003849:Deoxyguanosine / Q000378:metabolism; D003853:Deoxyribonucleosides / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011687:Purines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity
D003839:Deoxyadenosines; D003853:Deoxyribonucleosides; D011687:Purines; D011743:Pyrimidines; D003841:Deoxycytidine; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C020018:deoxyribonucleoside kinases; C021892:nucleoside phosphotransferase; D003849:Deoxyguanosine