Plasmodium falciparum MLH is schizont stage specific endonuclease

Tarique, Mohammed; Satsangi, Akash Tripathi; Ahmad, Moaz; Singh, Shailja; Tuteja, Renu

doi:10.1016/j.molbiopara.2011.10.012

Cited by 17 publications

(25 citation statements)

References 37 publications

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“…Helicase assay was performed using the previously characterized purified recombinant synthetic PfMLH [31], catalytically inactive N-terminal (PfMLHN), C-terminal (PfMLHC) [31] and the heat inactivated PfMLH and PfUDN. The strand displacement assay was used with both normal (substrate 1, Table S1) and direction-specific substrates (substrate 3B, Table S1).…”

Section: Methodsmentioning

confidence: 99%

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Plasmodium falciparum UvrD Helicase Translocates in 3′ to 5′ Direction, Colocalizes with MLH and Modulates Its Activity through Physical Interaction

et al. 2012

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Malaria is a global disease and a major health problem. The control of malaria is a daunting task due to the increasing drug resistance. Therefore, there is an urgent need to identify and characterize novel parasite specific drug targets. In the present study we report the biochemical characterization of parasite specific UvrD helicase from Plasmodium falciparum. The N-terminal fragment (PfUDN) containing UvrD helicase domain, which consists of helicase motifs Q, Ia–Id, II, III and most of motif IV, and the C-terminal fragment (PfUDC1) containing UvrD helicase C terminal domain, consisting of remaining part of motif IV and motifs IVa–IVc and 161 amino acids of intervening sequence between motif IV and V, possess ssDNA-dependent ATPase and DNA helicase activities in vitro. Using immunodepletion assays we show that the ATPase and helicase activities are attributable to PfUDN and PfUDC1 proteins. The helicase activity can utilize the hydrolysis of all the nucleotide and deoxynucleotide triphosphates and the direction of unwinding is 3′ to 5′. The endogenous P. falciparum UvrD contains the characteristic DNA helicase activity. PfUDN interacts with PfMLH (P. falciparum MutL homologue) and modulates the endonuclease activity of PfMLH and PfMLH positively regulates the unwinding activity of PfUDN. We show that PfUvrD is expressed in the nucleus distinctly in the schizont stages of the intraerythrocytic development of the parasite and it colocalizes with PfMLH. These studies will make an important contribution in understanding the nucleic acid transaction in the malaria parasite.

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Section: Methodsmentioning

confidence: 99%

“…Previously we have reported that the parasite P. falciparum genome contains the homologues of major components of MMR complex such as UvrD helicase and MutL homologue (MLH) [22]. In a recent study we have reported the isolation and characterization of MLH from P. falciparum [31].…”

Section: Introductionmentioning

confidence: 99%

Plasmodium falciparum UvrD Helicase Translocates in 3′ to 5′ Direction, Colocalizes with MLH and Modulates Its Activity through Physical Interaction

et al. 2012

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“…The pre-immune sera produced no detectable fluorescence (data not shown). To further confirm the stage-specific expression, the co-localization studies were performed with PfMLH, which was recently reported to be expressed specifically in schizont stages of the intraerythrocytic development in the parasite [42]. The slides were fixed and blocked in FBS as described above.…”

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confidence: 99%

“…After washing the slides were incubated with purified IgG of anti-PfRuvB3 antibodies at 1:200 dilutions as described above and washed. The slides were further incubated with purified IgG of anti-PfMLH [42] (raised in rabbit) at 1:200 dilutions in PBS containing 10% FBS for 1 h at 37 • C. After washing, the slides were further incubated for 1 h at 37 • C with appropriate secondary antibodies (Alexa 488 (green)-conjugated) anti-mouse IgG (Invitrogen) and (Alexa 594 (red)-conjugated) anti-rabbit IgG (Invitrogen) both diluted 1:500 in PBS containing 10% FBS. The slides were washed and mounted with antifade-DAPI reagent and viewed under oil immersion.…”

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Novel RuvB nuclear ATPase is specific to intraerythrocytic mitosis during schizogony of Plasmodium falciparum

Ahmad

Singh

Afrin

et al. 2012

Molecular and Biochemical Parasitology

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“…We have reported that the N-terminal and first half of the C-terminal of PfUvrD contain the characteristic helicase and ATPase activities and the direction of unwinding is 3′ to 5′ 10 . We have also shown that PfUvrD colocalizes with MLH (MutL homolog) and modulates its activity through physical interaction 10 , 11 . Furthermore the alignment with UvrD from E. coli showed the presence of all the conserved motifs in the UvrD of P. falciparum which suggests that PfUvrD is more homologous to prokaryotic system 10 …”

Section: Introductionmentioning

confidence: 99%

In silico analysis ofPlasmodiumspecies specific UvrD helicase

Tuteja

2013

Communicative & Integrative Biology

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Malaria is still a devastating disease caused by the mosquito-transmitted parasite Plasmodium, particularly Plasmodium falciparum. During the last few years the situation has worsened in many ways, mainly due to malarial parasites becoming increasingly resistant to several anti-malarial drugs. Thus there is an urgent need to find alternate ways to control malaria and therefore it is necessary to identify new drug targets and new classes of anti-malarial drugs. A malaria vaccine would be the ultimate weapon to fight this deadly disease but unfortunately despite encouraging advances a vaccine is not likely soon. DNA helicases from the PcrA/UvrD/Rep (PUR) subfamily are important for the survival of the various organisms, mainly pathogenic bacteria. Members from this subfamily can be targeted and inhibited by a variety of synthetic compounds. Using bioinformatics analysis we have shown that UvrD from this subfamily is the only member present in the P. falciparum genome, while PcrA and Rep are absent in the genome. UvrD from the parasite shows no homology to any protein or enzyme from human and thus can be considered as a strong potential drug target. In the present study we report an in silico analysis of this important enzyme from a variety of Plasmodium species. The results suggest that among all the species of Plasmodium, P. falciparum contains the largest UvrD and this enzyme is variable at the sequence and structural level.

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Plasmodium falciparum MLH is schizont stage specific endonuclease

Cited by 17 publications

References 37 publications

Plasmodium falciparum UvrD Helicase Translocates in 3′ to 5′ Direction, Colocalizes with MLH and Modulates Its Activity through Physical Interaction

Plasmodium falciparum UvrD Helicase Translocates in 3′ to 5′ Direction, Colocalizes with MLH and Modulates Its Activity through Physical Interaction

Novel RuvB nuclear ATPase is specific to intraerythrocytic mitosis during schizogony of Plasmodium falciparum

In silico analysis ofPlasmodiumspecies specific UvrD helicase

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