Mitochondrial apurinic/apyrimidinic endonuclease Apn1 is not critical for the completion of the Plasmodium berghei life cycle

Srivastava, Pratik Narain; Narwal, Sunil Kumar; Mishra, Satish

doi:10.1016/j.dnarep.2021.103078

Cited by 6 publications

(5 citation statements)

References 36 publications

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“…There is a proposition that microhomology-mediated end joining (alt-EJ) could also be an important DSB repair mechanism of the mtDNA in model organisms ( 19 , 20 ). In P. falciparum , two apurinic/apyrimidinic (AP) endonucleases have been found in the mitochondria ( 12 – 14 ), suggesting the existence of the BER mechanism in the parasite mitochondria. Even though the alt-EJ mechanism is discovered in this parasite ( 21 ), it is currently unknown whether this DSB repair mechanism contributes to the repair of mtDNA.…”

Section: Discussionmentioning

confidence: 99%

“…The mitochondrial DNA (mtDNA) replication is believed to take place either via the rolling-circle mode or by involving recombination intermediates ( 11 ). Among the DNA repair mechanisms, only the base excision repair (BER) mechanism is characterized in Plasmodium mitochondria ( 12 – 14 ). In model organisms, factors involved in mismatch repair (MMR) and nucleotide excision repair (NER) have been identified in the mitochondrial extracts ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

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Bloom Helicase Along with Recombinase Rad51 Repairs the Mitochondrial Genome of the Malaria Parasite

Jha

Gahlawat

Bhattacharyya

et al. 2021

mSphere

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Bloom Helicase Along with Recombinase Rad51 Repairs the Mitochondrial Genome of the Malaria Parasite

Jha

Gahlawat

Bhattacharyya

et al. 2021

mSphere

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“…The attempts to disrupt PbExo ( PB ANKA_0301700) were performed with a replacement plasmid containing GFP reporter and hDHFR:yFCU selection marker ( 56 ). Two 5′- and 3′-UTR homology regions F1 and F2 (0.57 Kb) were amplified using primers 1430/1431 and 1432/1433 and cloned at SalI and NotI/AscI respectively in the Pb C-GFP-hDHFR:yFCU vector.…”

Section: Methodsmentioning

confidence: 99%

A Plasmodium apicoplast-targeted unique exonuclease/FEN exhibits interspecies functional differences attributable to an insertion that alters DNA-binding

Chatterjee,

Tiwari,

Gupta

et al. 2024

Nucleic Acids Research

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The human malaria parasite Plasmodium falciparum genome is among the most A + T rich, with low complexity regions (LCRs) inserted in coding sequences including those for proteins targeted to its essential relict plastid (apicoplast). Replication of the apicoplast genome (plDNA), mediated by the atypical multifunctional DNA polymerase PfPrex, would require additional enzymatic functions for lagging strand processing. We identified an apicoplast-targeted, [4Fe–4S]-containing, FEN/Exo (PfExo) with a long LCR insertion and detected its interaction with PfPrex. Distinct from other known exonucleases across organisms, PfExo recognized a wide substrate range; it hydrolyzed 5′-flaps, processed dsDNA as a 5′-3′ exonuclease, and was a bipolar nuclease on ssDNA and RNA–DNA hybrids. Comparison with the rodent P. berghei ortholog PbExo, which lacked the insertion and [4Fe–4S], revealed interspecies functional differences. The insertion-deleted PfExoΔins behaved like PbExo with a limited substrate repertoire because of compromised DNA binding. Introduction of the PfExo insertion into PbExo led to gain of activities that the latter initially lacked. Knockout of PbExo indicated essentiality of the enzyme for survival. Our results demonstrate the presence of a novel apicoplast exonuclease with a functional LCR that diversifies substrate recognition, and identify it as the candidate flap-endonuclease and RNaseH required for plDNA replication and maintenance.

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“…To replace PbCmanT with a targeting construct, two homology regions (F1, 5′ UTR and F2, 3′ UTR) were amplified using primers 1292/1293 and 1294/1295 and cloned at SalI and NotI/AscI, respectively, in the pBC-GFP-hDHFR:yFCU vector as described previously. 14 The resulting construct was linearized with XhoI/AscI and transfected in MRA-311 as described previously to obtain the CmanT − parasite line. 21 Site specific integration PCRs were used to confirm gene knockout by using primers 1603/1225 and 1215/1604 for 5′ and 3′ integrations, respectively.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…PbCmanT gene was disrupted by double-crossover homologous recombination employing a replacement strategy, as described previously (Figure 1A). 14 Successful disruption of the CmanT gene was confirmed by diagnostic PCR and Southern blot (Figure 1B and C). The knockout strain was also complemented for function restoration (Figure 1D and E).…”

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confidence: 99%

C-Mannosyltransferase Is Essential for Malaria Transmission in Plasmodium berghei

et al. 2022

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C-Mannosylation of the thrombospondin type I repeat (TSR) domains is one of the most important factors involved in their function. It occurs on the first tryptophan of the WXXWXXC conserved motif where the tryptophan is usually surrounded by arginine or lysine forming the ligand-binding stretch of this sticky domain. It is found in its canonical or modified forms in many Plasmodium proteins. TSR containing proteins such as thrombospondin-like anonymous protein (TRAP), circumsporozoite protein (CSP), CSP and TRAP related protein (CTRP), and secreted protein with altered thrombospondin repeat (SPATR) have all been shown to be important for various parasite processes and life cycle stages. Here, we show that C-mannosylation catalyzing enzyme Cmannosyltransferase (CmanT) plays an essential role in malaria transmission in Plasmodium berghei. Disruption of the CmanT does not affect asexual blood stage propagation or gametocyte development but abolishes the formation of oocysts in mosquitoes. CmanT knockout (CmanT − ) parasites showed normal ookinete formation; however, these ookinetes failed in their ability to glide. CmanT − was complemented by reintroducing the gene, restoring mosquito transmission to wild-type level. We also investigated the effect of C-mannosylation on the folding and heparin-binding capacity of the Plasmodium falciparum TRAP TSR domain in silico, which suggested that this phenotype should be due to its involvement in the global stabilization of TSR residue side chain interactions.

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Mitochondrial apurinic/apyrimidinic endonuclease Apn1 is not critical for the completion of the Plasmodium berghei life cycle

Cited by 6 publications

References 36 publications

Bloom Helicase Along with Recombinase Rad51 Repairs the Mitochondrial Genome of the Malaria Parasite

Bloom Helicase Along with Recombinase Rad51 Repairs the Mitochondrial Genome of the Malaria Parasite

A Plasmodium apicoplast-targeted unique exonuclease/FEN exhibits interspecies functional differences attributable to an insertion that alters DNA-binding

C-Mannosyltransferase Is Essential for Malaria Transmission in Plasmodium berghei

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