Molecular Investigation of Genetic Signatures of Selection in </i>Plasmodium falciparum</i> Actin-Binding Protein Coronin, Cysteine Desulfurase, and Plasmepsin 2 Gene in Mbita Field Isolates, Western Kenya

Abstract

Background: Plasmodium falciparum (Pf) resistance to antimalarial drugs is a major impediment to malaria control. The Pf.Kelch 13 (PfK13) gene has been largely reported to be associated with artemisinin resistance. However, recent studies have shown artemisinin resistance without Kech13 mutations suggesting the implication of others genes in artemisinin resistance. In this current study, we focused on mutations in Pf.actin-binding protein coronin, Pf.cysteine desulfurase and Pf.plasmepsin 2 gene, three putative candidates recently were reported to be involved in artemisinin, lumefantrine and piperaquine resistance respectively. Method: Archived blood samples previously collected from asymptomatic school children from December 2016 to October 2018 were used in this study. Genomic DNA was extracted using ISOLATE II Genomic DNA kit. After PCR amplification, amplicons were purified and sequenced by capillary sequencing. Reads were analyzed for the identification of point mutations previously reported to be involved in drug selection. Results: Mutations R100K, and G50E involved in reduced artemisinin susceptibility were detected in Pfcoronin.

Artemisinin resistance mutations inPfcoroninimpede hemoglobin uptake

“…ART remains a crucial component of front-line antimalarial therapies, and its antimalarial efficacy must be protected. The current study underlines the potential threat of Pfcoronin -mediated resistance—in fact, the Pfcoronin R100K mutation was recently reported in Kenya, although artemisinin sensitivity/resistance was not assessed 45 . Other Pfcoronin polymorphisms were recently identified in Uganda 46 , and these polymorphisms were associated with ART resistance.…”

Prevalence of HIV-1 Natural Polymorphisms and Integrase-Resistance-Associated Mutations in African Children

“…Such frequencies can be attributed to natural genetic variability, frequently observed in the integrase gene [ 35 , 44 ]. The oscillation of the frequency of the unknown-effect polymorphism mutations may result from genetic drift [ 45 ] since HIV-1 is subject to genetic variability. Further investigations are required to elucidate the association of these unknown mutations with INI-based regimens.…”

Absolute quantification of gene copy number variation in Plasmodium falciparum novel putative genes associated to drug resistance (the actin-binding coronin protein, cysteine desulfurase and plasmepsin 2), using Eva green-based quantitative PCR

“…The protein is conserved, and is expressed in the merozoite's periphery and also localized in late schizogony [50]. The coronin protein contains β-propeller domain like PfKelch13 protein [31] and it processes WD40 repeats (repetitive residues composed of tryptophan (W) -aspartic acid (D) at the Cterminus while having glycine (G) -histidine (H) at the N terminus) spanning the β-propeller domain which serves protein-DNA and protein-protein interaction and is also involved in multiples functions such as chemical inhibitors pathways, regulation of transcription, signal transduction [51,52,41] Pfcoronin is also implicated in vesicular transport, cytoskeletal motility F-actin binding and its structural organisation [31]. These functions highlight the prominent role of Pfcoronin protein in the P.falciparum parasite.…”

“…The protein is involved in the Ironsulphur biosynthesis and its delivery to different metabolic pathways for instance the Fe-S complex which is modulated in apicomplexans during drug resistance, and stress conditions [32] In P.falciparum, the iron homeostasis is substantive for blood-stage parasite development and is implied in the mechanisms of quinolones drug, _ the lumefantrine drug family [32]. Moreover, many important features were predicted in the cysteine desulfurase protein including Aminotran_5 domain, disordered regions covering low complexity motifs [41] those functions include adhesion, signal transduction, molecular recognition, interaction with phospholipid bilayers, protein modi cation, and modulation of protein translation [40,54] Pfplasmepsin 2 gene was found to be the less polymorphic gene in copy number variation, in total 6 isolates showed copy number variation. However, the highest CNVs detected in this study was found in the Pfplasmepsin 2 gene, three isolates (OS062/17, KM019/18, and AL106/16) showed (9.63, 9.41 and 9.18 copies) of plasmepsin 2 gene respectively (Fig 5 .…”

“…A total of 36 samples (14 for Pfcysteine desulfurase, 13 for Pfpm2 and 9 for Pfcoronin) were assessed for copy number variation. These samples were selected based on their sequencing results that were found to harbour signi cant mutations among which some have been reported to be associated with ACT drug reduced susceptibility [41]. The ampli cation was carried out using Mic qPCR Cycler, v2.8.1 (Biomolecular systems, Australia).…”