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Enzyme Kinetics Of Dutpase From The Planarian Dugesia Ryukyuensis, Shahanoor Alam, Hideaki Moriyama
Enzyme Kinetics Of Dutpase From The Planarian Dugesia Ryukyuensis, Shahanoor Alam, Hideaki Moriyama
School of Biological Sciences: Faculty Publications
Objective: Planarians including Dugesia ryukyuensis (Dr) have strong regenerative abilities that require enhanced DNA replication. Knockdown of the DUT gene in Dr, which encodes deoxyuridine 5′-triphosphate pyrophosphatase (dUTPase), promotes DNA fragmentation, inhibits regeneration, and eventually leads to death. dUTPase catalyzes the hydrolysis of dUTP to dUMP and pyrophosphate. dUTPase is known to prevent uracil misincorporation in DNA by balancing the intracellular ratio between dUTP and dTTP, and contributes to genome stability. Nevertheless, the catalytic performance of Dr-dUTPase has not been reported.
Results: To confirm the catalytic activity of Dr-dUTPase, we cloned and expressed Dr-DUT in E. coli. Then, we purified …