'DEAD-box' helicase from Plasmodium falciparum is active at wide pH and is schizont stage-specific.
OBJECTIVE
DNA helicases catalyse unwinding of duplex DNA in an ATP-dependent manner and are involved in all the basic genetic processes. In order to study these important enzymes in the human malaria parasite we have recently cloned the first full-length 'DEAD-box' helicase gene from Plasmodium falciparum (3D7). In the present study, we report some of the important activities of the encoded protein.


METHODS
We have expressed the P. falciparum helicase in Escherichia coli and characterised the encoded biochemically active helicase protein. The characterisation of the protein was carried out using radioactively labeled substrate and the standard strand displacement assay. The localisation of the enzyme was studied using immunofluorescence assay.


CONCLUSIONS
P. falciparum helicase gene is 1551 bp in length and encodes for a protein consisting of 516 amino acid residues with a predicted molecular mass of 59.8 kDa. The protein is designated as Plasmodium falciparum DEAD-box helicase 60 kDa in size (PfDH60). Purified PfDH60 showed ATP and Mg2+ dependent DNA unwinding, ssDNA-dependent ATPase and ATP-binding activities. Interestingly, this is a unique helicase because it works at a wide pH range (from 5.0-10.0). The peak expression of PfDH60 is mainly in schizont stages of the development of P. falciparum, where DNA replication is active. The cell-cycle dependent expression suggests that PfDH60 may be involved in the process of DNA replication and distinct cellular processes in the parasite and this study should make an important contribution in our better understanding of DNA metabolic pathways in the parasite.
Pradhan
Arun
A
Malaria Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
Chauhan
Virander S
VS
Tuteja
Renu
R
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003001:Cloning, Molecular; D053487:DEAD-box RNA Helicases; D004265:DNA Helicases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004261:DNA Replication; D016054:DNA, Protozoan / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006863:Hydrogen-Ion Concentration; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D020365:RNA Helicases; D052939:Schizonts / Q000201:enzymology* / Q000254:growth & development*
D016054:DNA, Protozoan; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases