L-ATP is recognized by some cellular and viral enzymes: does chance drive enzymic enantioselectivity?
We demonstrate that l-ATP is recognized by some enzymes that are involved in the synthesis of nucleotides and nucleic acids. l-ATP, as well as its natural d-enantiomer, acts as a phosphate donor in the reaction catalysed by human deoxycytidine kinase, whereas it is not recognized by either enantioselective human thymidine kinase or non-enantioselective herpes virus thymidine kinase. l-ATP strongly inhibits (Ki 80 microM) the synthesis of RNA primers catalysed by DNA primase associated with human DNA polymerase alpha, whereas RNA synthesis catalysed by Escherichia coli RNA polymerase is completely unaffected. Moreover, l-ATP competitively inhibits ATP-dependent T4 DNA ligase (Ki 25 microM), suggesting that it interacts with the ATP-binding site of the enzyme. Kinetic studies demonstrated that l-ATP cannot be used as a cofactor in the ligase-catalysed joining reaction. On the other hand, l-AMP is used by T4 DNA ligase to catalyse the reverse reaction, even though a high level of intermediate circular nicked DNA molecules accumulates. Our results suggest that a lack of enantioselectivity of enzymes is more common than was believed a few years ago, and, given the absence of selective constraints against l-nucleosides in Nature, this may depend on chance more than on evolutionary strategy.
Verri
A
A
Istituto di Genetica Biochimica ed Evoluzionistica, CNR, Via Abbiategrasso 207, I-27100 Pavia, Italy.
Montecucco
A
A
Gosselin
G
G
Boudou
V
V
Imbach
J L
JL
Spadari
S
S
Focher
F
F
D000249:Adenosine Monophosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry*; D004247:DNA / Q000096:biosynthesis*; D011088:DNA Ligases / Q000737:chemistry; D019915:DNA Primase / Q000037:antagonists & inhibitors / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D003842:Deoxycytidine Kinase / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D018139:Simplexvirus / Q000201:enzymology; D013237:Stereoisomerism; D013937:Thymidine Kinase / Q000737:chemistry
D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D004247:DNA; D013937:Thymidine Kinase; D003842:Deoxycytidine Kinase; D019915:DNA Primase; D012321:DNA-Directed RNA Polymerases; D011088:DNA Ligases