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Ribozyme cleavage of Plasmodium falciparum gyrase A gene transcript affects the parasite growth

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Abstract

Deoxyribonucleic acid (DNA) gyrase is an important enzyme that facilitates the movement of replication and transcription complexes through DNA by creating negative supercoils ahead of the complex. Its presence in Plasmodium falciparum is now established and considered a good drug target since it is absent in the human host. The sequence of P. falciparum gyrase A subunit was analyzed for its messenger ribonucleic acid (mRNA) folding as well as target accessibility for ribozymes. The four GUC triplet sites identified at 334, 491, 1907, and 2642 nucleotide positions of the Gyrase A mRNA were also accessible to oligos by RNase H assay. Site GUC491 was optimally accessible followed by GUC1907, GUC334, and GUC2642 sites. Ribozymes were produced against all these sites and tested for their in vitro transcript cleavage potentials where RZ491 showed the maximum cleavage rate. Therefore, this ribozyme (RZ491) was chemically synthesized albeit with modifications so as to make it resistant against ribonuclease attack. The modified ribozyme retained its cleavage potential and was able to inhibit the P. falciparum parasite growth up to 49.54% and 74.77% at 20 and 30 μM ribozyme concentrations, respectively, as compared to the untreated culture. However, up to 20% and 24.32% parasite growth inhibition was observed at the same ribozyme concentrations of 20 and 30 μM when compared with control ribozyme-treated cultures. This ribozyme as well as other targets identified here can be investigated further to develop the effective chemotherapeutic agents against malaria.

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Acknowledgments

We thank Dr. Sukla Biswas and Dr. C.R. Pillai for their help in parasite culture. Senior Research Fellowships (to AA) from Council of Scientific and Industrial Research (CSIR) is highly acknowledged. We thank Dr. P. K. Yadava for useful discussions regarding RNA folding and ribozyme designing. The Biotechnology Information System (BTIS) facility is gratefully acknowledged.

Funding

Financial supports for the work were granted by Department of Biotechnology (DBT) and ICMR, Government of India.

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  1. Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Anwar Ahmed & Yagya D. Sharma

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  1. Anwar Ahmed
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  2. Yagya D. Sharma
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Correspondence to Yagya D. Sharma.

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Ahmed, A., Sharma, Y.D. Ribozyme cleavage of Plasmodium falciparum gyrase A gene transcript affects the parasite growth. Parasitol Res 103, 751–763 (2008). https://doi.org/10.1007/s00436-008-1036-y

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