Inhibition of unwinding and ATPase activities of<i>Plasmodium falciparum</i>Dbp5/DDX19 homologue

Mehta, Jatin; Tuteja, Renu

doi:10.4161/cib.4.3.14778

Cited by 7 publications

(6 citation statements)

References 21 publications

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“…PfRuvB3 is more sensitive to daunorubicin (IC 50 of 0.76 μM) than other helicases, such as P. falciparum DNA helicase A (PfDH A) [22], P. falciparum UvrD helicase (PfUDN) [40], P. falciparum 45 kDa helicase (PfH45) [41], P. falciparum 60 kDa DNA helicase (PfDH60) [42] and human DNA helicase (HDH II) [43] (IC 50s of 2.0, 4.4, 5.0 3.0, 6.23 μM, respectively). However, it is less sensitive to netropsin (IC 50 of 7.09 μM) than PfUDN, PfH45, PfDH60 and P. falciparum Dbp5/DDX19 homolog (PfD66) [44] (IC 50s of 3.3, 0.8, 0.5, 3.2 μM, respectively). Inhibition of PfRuvB3 activity by daunorubicin (IC 50 of 0.76 μM) and doxorubicin (IC 50 of 2.6 μM) did not correlate with parasite growth inhibition (IC 50s of 2.5 and 1.5 μM respectively) [22] suggesting that they may have different cell permeability and metabolic properties.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3

Limudomporn

Moonsom

Leartsakulpanich

et al. 2016

Malar J

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BackgroundMalaria is one of the most serious and widespread parasitic diseases affecting humans. Because of the spread of resistance in both parasites and the mosquito vectors to anti-malarial drugs and insecticides, controlling the spread of malaria is becoming difficult. Thus, identifying new drug targets is urgently needed. Helicases play key roles in a wide range of cellular activities involving DNA and RNA transactions, making them attractive anti-malarial drug targets.MethodsATP-dependent DNA helicase gene (PfRuvB3) of Plasmodium falciparum strain K1, a chloroquine and pyrimethamine-resistant strain, was inserted into pQE-TriSystem His-Strep 2 vector, heterologously expressed and affinity purified. Identity of recombinant PfRuvB3 was confirmed by western blotting coupled with tandem mass spectrometry. Helicase and ATPase activities were characterized as well as co-factors required for optimal function.ResultsRecombinant PfRuvB3 has molecular size of 59 kDa, showing both DNA helicase and ATPase activities. Its helicase activity is dependent on divalent cations (Cu2+, Mg2+, Ni+2 or Zn+2) and ATP or dATP but is inhibited by high NaCl concentration (>100 mM). PfPuvB3 is unable to act on blunt-ended duplex DNA, but manifests ATPase activity in the presence of either single- or double-stranded DNA. PfRuvB3.is inhibited by doxorubicin, daunorubicin and netropsin, known DNA helicase inhibitors.ConclusionsPurified recombinant PfRuvB3 contains both DNA helicase and ATPase activities. Differences in properties of RuvB between the malaria parasite obtained from the study and human host provide an avenue leading to the development of novel drugs targeting specifically the malaria form of RuvB family of DNA helicases.

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Section: Discussionmentioning

confidence: 99%

Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3

Limudomporn

Moonsom

Leartsakulpanich

et al. 2016

Malar J

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“…These compounds can be natural or synthetic aminoglycosides and nonaminoglycosides [100,[102][103][104][105][106]. Interestingly, several compounds have already been identified to inhibit the helicase and ATPase activities of the Plasmodium falciparum Dbp5/DDX19 homolog PfD66 [107]. Although promising, a great concern with these compounds is that they lack drug target specificity.…”

Section: Termination Readthrough and Nonsense Mediated Decay (Nmd)mentioning

confidence: 99%

Dbp5/DDX19 between Translational Readthrough and Nonsense Mediated Decay

Beißel

Große

Krebber

2020

IJMS

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The DEAD-box protein Dbp5 (human DDX19) remodels RNA-protein complexes. Dbp5 functions in ribonucleoprotein export and translation termination. Termination occurs, when the ribosome has reached a stop codon through the Dbp5 mediated delivery of the eukaryotic termination factor eRF1. eRF1 contacts eRF3 upon dissociation of Dbp5, resulting in polypeptide chain release and subsequent ribosomal subunit splitting. Mutations in DBP5 lead to stop codon readthrough, because the eRF1 and eRF3 interaction is not controlled and occurs prematurely. This identifies Dbp5/DDX19 as a possible potent drug target for nonsense suppression therapy. Neurodegenerative diseases and cancer are caused in many cases by the loss of a gene product, because its mRNA contained a premature termination codon (PTC) and is thus eliminated through the nonsense mediated decay (NMD) pathway, which is described in the second half of this review. We discuss translation termination and NMD in the light of Dbp5/DDX19 and subsequently speculate on reducing Dbp5/DDX19 activity to allow readthrough of the PTC and production of a full-length protein to detract the RNA from NMD as a possible treatment for diseases.

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“…PfD68 is also expressed in all the intraerythrocytic developmental stages (i.e., the ring, the trophozoite, and the schizont) of P. falciparum 3D7 strain and the localization of PfD68 was mainly in the cytoplasm, indicating that it plays most likely a role in cytoplasmic processes involving RNA. The DNA helicase and ATPase activities of PfD68 were reported to be inhibited by DNA‐binding compounds such as ethidium bromide, actinomycin, daunorubicin, DAPI, and netropsin . The intercalation of the compounds into the duplex DNA substrate probably blocks the sustained translocation of the enzyme that, in turn, inhibits the unwinding of the duplex substrate .…”

Section: Dead‐box Helicases Of Plasmodium Falciparummentioning

confidence: 99%

“…The DNA helicase and ATPase activities of PfD68 were reported to be inhibited by DNA‐binding compounds such as ethidium bromide, actinomycin, daunorubicin, DAPI, and netropsin . The intercalation of the compounds into the duplex DNA substrate probably blocks the sustained translocation of the enzyme that, in turn, inhibits the unwinding of the duplex substrate .…”

Section: Dead‐box Helicases Of Plasmodium Falciparummentioning

confidence: 99%

Unraveling the importance of the malaria parasite helicases

Tuteja

2017

The FEBS Journal

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Malaria is a human parasitic disease caused by infection from Plasmodium species, particularly Plasmodium falciparum. Each year millions of people are infected with malaria and large numbers of deaths result due to this deadly infection. P. falciparum contains 14 chromosomes, nearly 5400 genes and a multistage life cycle in humans and mosquitoes. The control of malaria is still a challenge as the parasite is continuously developing resistance to available antimalarial drugs and the mosquito vector is developing resistance to insecticides. The availability of P. falciparum genome has resulted in the identification of parasite‐specific proteins that can be targeted without harmful effects to the human host. Toward this goal, we have been working on the identification and characterization of helicases in order to find parasite‐specific helicases, which can be used as novel drug targets to tackle the rising problem of drug resistance. Helicases are ATP‐dependent nucleic acid unwinding enzymes. The P. falciparum genome analysis depicts that it contains some parasite‐specific helicases and homologs to most of the human helicases. Here, we present an overview of P. falciparum helicases and their importance in parasite growth and survival.

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Inhibition of unwinding and ATPase activities ofPlasmodium falciparumDbp5/DDX19 homologue

Cited by 7 publications

References 21 publications

Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3

Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3

Dbp5/DDX19 between Translational Readthrough and Nonsense Mediated Decay

Unraveling the importance of the malaria parasite helicases

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