Characterization of replication fork and phosphorylation stimulated Plasmodium falciparum helicase 45.
Helicases are essential enzymes, which play important role in the metabolism of nucleic acids. In the present study we report further characterization of PfH45 (Plasmodium falciparum helicase 45), which is an essential enzyme for parasite survival. The results show that the helicase activity of PfH45 is significantly stimulated by replication fork like structure. The studies using truncated derivatives of PfH45 show that its nucleic acid dependent ATPase activity resides in the N-terminal one third of the protein and its RNA and DNA-binding activity predominantly resides in the C-terminal two third of the protein. The phosphorylation of PfH45 by protein kinase C at Ser and Thr residues stimulated its DNA and RNA helicase and ssDNA and RNA-dependent ATPase activities. DNA-interacting compounds actinomycin, DAPI, daunorubicin, ethidium bromide, netropsin and nogalamycin were able to inhibit the helicase and ssDNA-dependent ATPase activity with apparent IC50 values ranging from 0.5 to 5.0 microM respectively. These compounds distinctively inhibit the helicase activity probably by forming complex with DNA and obstructing enzyme movement.
Malaria Group, International Centre for Genetic Engineering and Biotechnology, P. O. Box 10504, Aruna Asaf Ali Marg, New Delhi-110067, India.
Arun
A
Pradhan
Ejaz M
EM
Hussain
Renu
R
Tuteja
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism*; D004261:DNA Replication; D009690:Nucleic Acid Conformation; D010766:Phosphorylation; D010963:Plasmodium falciparum / Q000201:enzymology*; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism
D015800:Protozoan Proteins; D016601:RNA-Binding Proteins; D000251:Adenosine Triphosphatases; D004265:DNA Helicases