Functional role of lysine 12 in Leishmania major AQP1

Sharma, Meenakshi; Goutam, Mandal; Srotoswati, Mandal; Bhattacharjee, Hiranmoy; Mukhopadhyay, Rita

doi:10.1016/j.molbiopara.2015.07.005

Cited by 8 publications

(15 citation statements)

References 26 publications

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“…Oligomerization correlates with bidirectional glycerol flow but also pentamidine sensitivity as C-terminal tagged forms form oligomers with low efficiency and have poor transport activity. Furthermore, TbAQP2 is ubiquitylated and targeted to the lysosome, similar to mammalian AQPs [62].…”

Section: Discussionmentioning

confidence: 99%

Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance inTrypanosoma brucei

Quintana

Bueren-Calabuig

Zuccotto

et al. 2020

Preprint

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34Defining mode of action is vital for both developing new drugs and predicting 35 potential resistance mechanisms. African trypanosome pentamidine and 36 melarsoprol sensitivity is predominantly mediated by aquaglyceroporin 2 (TbAQP2), a 37 channel associated with water/glycerol transport. TbAQP2 is expressed at the flagellar 38 pocket membrane and chimerisation with TbAQP3 renders parasites resistant to both 39 drugs. Two models for how TbAQP2 mediates pentamidine sensitivity have emerged; 40 that TbAQP2 mediates pentamidine translocation or via binding to TbAQP2, with 41 subsequent endocytosis, but trafficking and regulation of TbAQPs is uncharacterised. 42We demonstrate that TbAQP2 is organised as a high order complex, is ubiquitylated 43 and transported to the lysosome. Unexpectedly, mutation of potential ubiquitin 44 conjugation sites, i.e. cytoplasmic lysine residues, reduced folding and tetramerization 45 efficiency and triggered ER retention. Moreover, TbAQP2/TbAQP3 chimerisation also 46 leads to impaired oligomerisation, mislocalisation, and increased turnover. These data 47suggest that TbAQP2 stability is highly sensitive to mutation and contributes towards 48 emergence of drug resistance. 49 50 Human African trypanosomiasis (HAT) is a neglected tropical disease affecting 52 sub-Saharan countries [1-4]. HAT progresses by two stages: a haemolymphatic 53 stage, in which the parasite successfully colonises the bloodstream, lymphatics, skin, 54 adipose tissue and organs and a meningoencephalic stage characterised by the 55 emergence of parasites in the central nervous system (CNS) [2,5]. Several drugs are 56 used to treat HAT; currently suramin and pentamidine are the drugs of choice for 57 treatment of the haemolymphatic stage of T. brucei rhodesiense and T. brucei 58 gambiense infections respectively, whereas melarsoprol, eflornithine or combined 59 nifurtimox-eflornithine (NECT) therapy are recommended for the meningoencephalic 60 stage [6,7]. 61 Two new drugs, fexinidazole and acoziborole, recently completed clinical trials 62 and opened a new front in HAT chemotherapy [8,9]. Drug development, successful 63 public health initiatives and active case-monitoring programs have all contributed to 64 the anticipated eradication of gambiense HAT as a major public health problem in the 65 coming decade [10]. However, vigilance and understanding of drug mechanisms and 66 possible resistance pathways remain essential to maintain this situation, and 67rhodesiense HAT cannot be eliminated in this way as it is highly zoonotic [11]. 68Genome-wide RNAi screens identified a number of genes associated with 69 pentamidine sensitivity that, together with evidence from melarsoprol-pentamidine 70 cross-resistance (MPXR), identified aquaglyceroporin 2 as the primary determinant for 71 drug-uptake [12,13], alongside lesser roles for the TbAT1/P2 aminopurine transporter 72 and the Low Affinity Pentamidine transporter LAPT1 [14]. 73 Aquaglyceroporins (AQPs) are an ancient family of multi-pass membrane 74 proteins, containing b...

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Section: Discussionmentioning

confidence: 99%

Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance inTrypanosoma brucei

Quintana

Bueren-Calabuig

Zuccotto

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…AQPs form homotetrameric complexes to transport water and low molecular weight solutes [14][15][16]. Independent expansions of AQP paralogues have served to diversify AQP function, and in mammals and Leishmania major both ubiquitylation and phosphorylation are important in modulating turnover and hence activity [69][70][71][72][73][74]. Significantly, the three trypanosome AQP paralogs derive from a single ancestral gene shared with Leishmania spp., and thus relative functions of paralogs are likely differentially distributed between major lineages.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance in Trypanosoma brucei

et al. 2020

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Defining mode of action is vital for both developing new drugs and predicting potential resistance mechanisms. Sensitivity of African trypanosomes to pentamidine and melarsoprol is predominantly mediated by aquaglyceroporin 2 (TbAQP2), a channel associated with water/ glycerol transport. TbAQP2 is expressed at the flagellar pocket membrane and chimerisation with TbAQP3 renders parasites resistant to both drugs. Two models for how TbAQP2 mediates pentamidine sensitivity have emerged; that TbAQP2 mediates pentamidine translocation across the plasma membrane or via binding to TbAQP2, with subsequent endocytosis and presumably transport across the endosomal/lysosomal membrane, but as trafficking and regulation of TbAQPs is uncharacterised this remains unresolved. We demonstrate that TbAQP2 is organised as a high order complex, is ubiquitylated and is transported to the lysosome. Unexpectedly, mutation of potential ubiquitin conjugation sites, i.e. cytoplasmicoriented lysine residues, reduced folding and tetramerization efficiency and triggered ER retention. Moreover, TbAQP2/TbAQP3 chimerisation, as observed in pentamidine-resistant parasites, also leads to impaired oligomerisation, mislocalisation and increased turnover. These data suggest that TbAQP2 stability is highly sensitive to mutation and that instability contributes towards the emergence of drug resistance.

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“…The role of MAP2 for phosphorylation of threonine for providing stability of LmAQP1 and then overexpression needs more attention with the goal of developing a new approach for drug targeting. On the other hand, development of some compounds that target MAP2 for preventing phosphorylation and increasing the half-life of AQP1 may be dangerous because MAP kinases in all organisms are familiar and therefore they might target the ones from human, too (18). Although AQP1 protein belongs to the vast family of AQP in all organisms but finding some point nucleotides inside it might be the better target for designing the drug.…”

Section: Discussionmentioning

confidence: 99%

Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Iso¬lates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran

Alijani

Hosseini

Ahmadian

et al. 2019

JAD

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Background: Regarding the antimonial-resistant of Leishmania spp., understanding of related mechanism is necessary. One of the most important involved molecules is aquaglyceropin1 (AQP1). The aim of this study was molecular analysis of AQP1 gene from antimonial-resistant clinical isolates and its expression. Methods: Overall, 150 patients with cutaneous leishmaniasis referring to the reference laboratories of Yazd and Varzaneh,, located 105km southeast of Isfahan and 240km away from Yazd, were assessed from Jun 2015 to Dec 2017. After sampling, staining was done and evaluated for Leishman by microscope. Samples were collected in RNAlater solution for gene expression analysis in non-healing isolates. DNA extraction was performed from each slide with Leishman body. All patients with L. major isolates detected by ITS1-PCR-RFLP were followed for finding the resistant isolates, consequence of molecular characterization of AQP1 using PCR-RFLP. Gene expression of AQP1 from all resistant isolates was assessed in comparison with the one in a sensitive isolate. Statistical analysis was done using SPSS. The significance level was considered ≤0.05. Results: Five isolates were detected as antimonial resistant. Molecular detection and identification were appeared that all were L. major. The molecular characterization of AQP1 showed G562A mutation. Gene expression of AQP1 in resistant isolates showed 1.67 fold higher than the sensitive isolate. Conclusion: We reported a new point mutation of G562A in AQP1 gene involved in molecular mechanism in resistant isolates.

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Functional role of lysine 12 in Leishmania major AQP1

Cited by 8 publications

References 26 publications

Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance inTrypanosoma brucei

Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance inTrypanosoma brucei

Instability of aquaglyceroporin (AQP) 2 contributes to drug resistance in Trypanosoma brucei

Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Iso¬lates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran

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