scholarly journals A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

2021 ◽  
Author(s):  
Zhenke Yang ◽  
Yang Shi ◽  
Huiting Cui ◽  
Shuzhen Yang ◽  
Han Gao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Bilayers ◽  
Malaria Parasite ◽  
Parasite Infection ◽  
Midgut Epithelium ◽  
Rodent Malaria Parasite ◽  
Membrane Asymmetry ◽  
Type Iv ◽  
Parasite Survival ◽  
Phospholipid Flippase

Mosquito midgut epithelium traversal is an essential component of transmission of malaria parasites. Phospholipid flippases are eukaryotic type IV ATPases (P4-ATPases), which in association with CDC50 cofactors, translocate phospholipids across lipid bilayers to maintain the membrane asymmetry. In this study, we investigated the function of a putative P4-ATPase, ATP7, from the rodent malaria parasite P. yoelii. Disruption of ATP7 results in block of parasite infection of mosquitoes. ATP7 is localized on the ookinete plasma membrane. While ATP7-depleted ookinetes are motile and capable of invading the midgut, they are quickly eliminated within the epithelial cells by a process that is independent from the mosquito complement-like immunity. ATP7 colocalizes and interacts with the flippase co-factor CDC50C. Importantly, depletion of CDC50C phenocopies ATP7 deficiency. ATP7-depleted ookinetes fail to translocate phosphatidylcholine (PC) across the plasma membrane, resulting in PC exposure at the ookinete surface. Lastly, ookinete microinjection into the mosquito hemocoel reverses the ATP7 deficiency phenotype. Our study identifies Plasmodium flippase as a novel mechanism of parasite survival in the midgut epithelium that is required for mosquito transmission.

scholarly journals A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

2021 ◽  
Vol 7 (30) ◽  
pp. eabf6015
Author(s):  
Zhenke Yang ◽  
Yang Shi ◽  
Huiting Cui ◽  
Shuzhen Yang ◽  
Han Gao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Bilayers ◽  
Malaria Parasite ◽  
Membrane Lipid ◽  
Plasmodium Yoelii ◽  
Midgut Epithelium ◽  
Rodent Malaria Parasite ◽  
Adenosine Triphosphatases ◽  
Parasite Survival ◽  
P Type

Mosquito midgut epithelium traversal is essential for malaria parasite transmission. Phospholipid flippases are eukaryotic type 4 P-type adenosine triphosphatases (P4-ATPases), which, in association with CDC50, translocate phospholipids across the membrane lipid bilayers. In this study, we investigated the function of a putative P4-ATPase, ATP7, from the rodent malaria parasite Plasmodium yoelii. Disruption of ATP7 blocks the parasite infection of mosquitoes. ATP7 is localized on the ookinete plasma membrane. While ATP7-depleted ookinetes are capable of invading the midgut, they are eliminated within the epithelial cells by a process independent from the mosquito complement-like immunity. ATP7 colocalizes and interacts with the flippase cofactor CDC50C. Depletion of CDC50C phenocopies ATP7 deficiency. ATP7-depleted ookinetes fail to uptake phosphatidylcholine across the plasma membrane. Ookinete microinjection into the mosquito hemocoel reverses the ATP7 deficiency phenotype. Our study identifies Plasmodium flippase as a mechanism of parasite survival in the midgut epithelium that is required for mosquito transmission.

scholarly journals Phospholipid flippase activities and substrate specificities of human type IV P-type ATPases localized to the plasma membrane.

2016 ◽  
Vol 291 (41) ◽  
pp. 21421-21421 ◽  
Author(s):  
Hiroyuki Takatsu ◽  
Gaku Tanaka ◽  
Katsumori Segawa ◽  
Jun Suzuki ◽  
Shigekazu Nagata ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Substrate Specificities ◽  
Human Type ◽  
Type Iv ◽  
P Type ◽  
Phospholipid Flippase

scholarly journals The PQ-loop protein Any1 segregates Drs2 and Neo1 functions required for viability and plasma membrane phospholipid asymmetry

2019 ◽  
Vol 60 (5) ◽  
pp. 1032-1042 ◽  
Author(s):  
Mehmet Takar ◽  
Yannan Huang ◽  
Todd R. Graham
Keyword(s):  
Plasma Membrane ◽  
Protein Interactions ◽  
Critical Role ◽  
Protein Protein Interactions ◽  
Membrane Asymmetry ◽  
Growth Defects ◽  
Type Iv ◽  
Organizational Feature ◽  
Membrane Type ◽  
P Type

Membrane asymmetry is a key organizational feature of the plasma membrane. Type IV P-type ATPases (P4-ATPases) are phospholipid flippases that establish membrane asymmetry by translocating phospholipids, such as phosphatidylserine (PS) and phospatidylethanolamine, from the exofacial leaflet to the cytosolic leaflet. Saccharomyces cerevisiae expresses five P4-ATPases: Drs2, Neo1, Dnf1, Dnf2, and Dnf3. The inactivation of Neo1 is lethal, suggesting Neo1 mediates an essential function not exerted by the other P4-ATPases. However, the disruption of ANY1, which encodes a PQ-loop membrane protein, allows the growth of neo1Δ and reveals functional redundancy between Golgi-localized Neo1 and Drs2. Here we show Drs2 PS flippase activity is required to support neo1Δ any1Δ viability. Additionally, a Dnf1 variant with enhanced PS flipping ability can replace Drs2 and Neo1 function in any1Δ cells. any1Δ also suppresses drs2Δ growth defects but not the loss of membrane asymmetry. Any1 overexpression perturbs the growth of cells but does not disrupt membrane asymmetry. Any1 coimmunoprecipitates with Neo1, an association prevented by the Any1-inactivating mutation D84G. These results indicate a critical role for PS flippase activity in Golgi membranes to sustain viability and suggests Any1 regulates Golgi membrane remodeling through protein-protein interactions rather than a previously proposed scramblase activity.

scholarly journals Exofacial membrane composition and lipid metabolism regulates plasma membrane P4-ATPase substrate specificity

2020 ◽  
Author(s):  
Bartholomew P. Roland ◽  
Bhawik K. Jain ◽  
Todd R. Graham
Keyword(s):  
Plasma Membrane ◽  
Membrane Lipids ◽  
Asymmetric Distribution ◽  
Membrane Composition ◽  
Membrane Biology ◽  
Membrane Asymmetry ◽  
Type Iv ◽  
Lipid Species ◽  
Endogenous Lipid ◽  
A Cell

AbstractThe plasma membrane of a cell is characterized by an asymmetric distribution of lipid species across the exofacial and cytofacial aspects of the bilayer. The regulation of membrane asymmetry is a fundamental characteristic of membrane biology, and is crucial for signal transduction, vesicle transport, and cell division. The type-IV family of P-ATPases, or P4-ATPases, establish membrane asymmetry by selection and transfer of a subset of membrane lipids from the lumenal or exofacial leaflet to the cytofacial aspect of the bilayer. It is still unclear how these enzymes sort through the spectrum of lipids within the membrane to identify their desired substrate(s) and how the membrane environment modulates this activity. Therefore, we tested how the yeast plasma membrane P4-ATPase, Dnf2, responds to changes in membrane composition induced by perturbation of endogenous lipid biosynthetic pathways or exogenous application of lipid. The primary substrates of Dnf2 are two chemically divergent lipids, glucosylceramide (GlcCer) and phosphatidylcholine ((PC) or their lyso-lipid derivatives), and we find that these substrates compete with each other for transport. Acutely inhibiting sphingolipid synthesis using myriocin attenuates transport of exogenously applied GlcCer without perturbing PC transport. Deletion of genes controlling later steps of glycosphingolipid production also perturb GlcCer transport to a greater extent than PC transport. Surprisingly, application of lipids that are poor transport substrates differentially affect PC and GlcCer transport by Dnf2, thus altering substrate preference. Our data indicate that Dnf2 exhibits exquisite sensitivity to the membrane composition; thus, providing feedback onto the function of the P4-ATPases.

scholarly journals Super-elasticity of plasma- and synthetic membranes by coupling of membrane asymmetry and liquid-liquid phase separation

2020 ◽  
Author(s):  
Jan Steinkühler ◽  
Tripta Bhatia ◽  
Ziliang Zhao ◽  
Reinhard Lipowsky ◽  
Rumiana Dimova
Keyword(s):  
Plasma Membrane ◽  
Phase Separation ◽  
Liquid Phase ◽  
Lipid Bilayers ◽  
Lipid Vesicles ◽  
Liquid Phase Separation ◽  
Elastic Response ◽  
Plasma Membrane Vesicles ◽  
Membrane Asymmetry ◽  
Liquid Liquid Phase Separation

AbstractBiological cells are contained by a fluid lipid bilayer (plasma membrane, PM) that allows for large deformations, often exceeding 50% of the initial (or projected) PM area. Biochemically isolated lipids self-organize into membranes, but the extraordinary deformability of the plasma membrane is lost. Pure lipid bilayers are prone to rupture at small (<2-4%) area strains and this limits progress for synthetic reconstitution of cellular features such as migration, phagocytosis and division. Here, we show that by preserving PM structure and composition during isolation from cells, vesicles with cell-like elasticity are obtained. We found that these plasma membrane vesicles store significant area in the form of nanotubes in their lumen. These are recruited by mechanical tension applied to the outer vesicle membrane showing an apparent elastic response. This “super-elastic” response emerges from the interplay of lipid liquid-liquid phase separation and membrane asymmetry. This finding allows for bottom-up engineering of synthetic vesicles that appear over one magnitude softer and with three fold larger deformability than conventional lipid vesicles.

scholarly journals Phospholipid Flippase Activities and Substrate Specificities of Human Type IV P-type ATPases Localized to the Plasma Membrane

2014 ◽  
Vol 289 (48) ◽  
pp. 33543-33556 ◽  
Author(s):  
Hiroyuki Takatsu ◽  
Gaku Tanaka ◽  
Katsumori Segawa ◽  
Jun Suzuki ◽  
Shigekazu Nagata ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Substrate Specificities ◽  
Human Type ◽  
Type Iv ◽  
P Type ◽  
Phospholipid Flippase

scholarly journals The phospholipid flippase ATP9A is required for the recycling pathway from the endosomes to the plasma membrane

2016 ◽  
Vol 27 (24) ◽  
pp. 3883-3893 ◽  
Author(s):  
Yoshiki Tanaka ◽  
Natsuki Ono ◽  
Takahiro Shima ◽  
Gaku Tanaka ◽  
Yohei Katoh ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Bilayers ◽  
Golgi Complex ◽  
Membrane Trafficking ◽  
Glucose Transporter ◽  
Glucose Transporter 1 ◽  
Recycling Endosomes ◽  
Type Iv ◽  
Late Endosomes ◽  
P Type

Type IV P-type ATPases (P4-ATPases) are phospholipid flippases that translocate phospholipids from the exoplasmic (or luminal) to the cytoplasmic leaflet of lipid bilayers. In Saccharomyces cerevisiae, P4-ATPases are localized to specific subcellular compartments and play roles in compartment-mediated membrane trafficking; however, roles of mammalian P4-ATPases in membrane trafficking are poorly understood. We previously reported that ATP9A, one of 14 human P4-ATPases, is localized to endosomal compartments and the Golgi complex. In this study, we found that ATP9A is localized to phosphatidylserine (PS)-positive early and recycling endosomes, but not late endosomes, in HeLa cells. Depletion of ATP9A delayed the recycling of transferrin from endosomes to the plasma membrane, although it did not affect the morphology of endosomal structures. Moreover, depletion of ATP9A caused accumulation of glucose transporter 1 in endosomes, probably by inhibiting their recycling. By contrast, depletion of ATP9A affected neither the early/late endosomal transport and degradation of epidermal growth factor (EGF) nor the transport of Shiga toxin B fragment from early/recycling endosomes to the Golgi complex. Therefore ATP9A plays a crucial role in recycling from endosomes to the plasma membrane.

scholarly journals Inhibitors of Eicosanoid Biosynthesis Reveal that Multiple Lipid Signaling Pathways Influence Malaria Parasite Survival in Anopheles gambiae

Insects ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 307 ◽  
Author(s):  
Kwon ◽  
Smith
Keyword(s):  
Fatty Acids ◽  
Anopheles Gambiae ◽  
Malaria Parasite ◽  
Parasite Infection ◽  
Immune Homeostasis ◽  
Dodecanoic Acid ◽  
Parasite Survival ◽  
Mosquito Anopheles Gambiae ◽  
Inflammatory Drugs ◽  
Specific Inhibitors

Eicosanoids are bioactive signaling lipids derived from the oxidation of fatty acids that act as important regulators of immune homeostasis and inflammation. As a result, effective anti-inflammatory drugs have been widely used to reduce pain and inflammation which target key eicosanoid biosynthesis enzymes. Conserved from vertebrates to insects, the use of these eicosanoid pathway inhibitors offer opportunities to evaluate the roles of eicosanoids in less-characterized insect systems. In this study, we examine the potential roles of eicosanoids on malaria parasite survival in the mosquito Anopheles gambiae. Using Plasmodium oocyst numbers to evaluate parasite infection, general or specific inhibitors of eicosanoid biosynthesis pathways were evaluated. Following the administration of dexamethasone and indomethacin, respective inhibitors of phospholipid A2 (PLA2) and cyclooxygenase (COX), oocyst numbers were unaffected. However, inhibition of lipoxygenase (LOX) activity through the use of esculetin significantly increased oocyst survival. In contrast, 12-[[(tricyclo[3.3.1.13,7]dec-1-ylamino)carbonyl]amino]-dodecanoic acid (AUDA), an inhibitor of epoxide hydroxylase (EH), decreased oocyst numbers. These experiments were further validated through RNAi experiments to silence candidate genes homologous to EH in An. gambiae to confirm their contributions to Plasmodium development. Similar to the results of AUDA treatment, the silencing of EH significantly reduced oocyst numbers. These results imply that specific eicosanoids in An. gambiae can have either agonist or antagonistic roles on malaria parasite survival in the mosquito host.

scholarly journals Exofacial membrane composition and lipid metabolism regulates plasma membrane P4-ATPase substrate specificity

2020 ◽  
Vol 295 (52) ◽  
pp. 17997-18009 ◽  
Author(s):  
Bhawik Kumar Jain ◽  
Bartholomew P. Roland ◽  
Todd R. Graham
Keyword(s):  
Plasma Membrane ◽  
Membrane Lipids ◽  
Ergosterol Biosynthesis ◽  
Asymmetric Distribution ◽  
Membrane Composition ◽  
Membrane Biology ◽  
Membrane Asymmetry ◽  
Type Iv ◽  
Lipid Species ◽  
A Cell

The plasma membrane of a cell is characterized by an asymmetric distribution of lipid species across the exofacial and cytofacial aspects of the bilayer. Regulation of membrane asymmetry is a fundamental characteristic of membrane biology and is crucial for signal transduction, vesicle transport, and cell division. The type IV family of P-ATPases, or P4-ATPases, establishes membrane asymmetry by selection and transfer of a subset of membrane lipids from the lumenal or exofacial leaflet to the cytofacial aspect of the bilayer. It is unclear how P4-ATPases sort through the spectrum of membrane lipids to identify their desired substrate(s) and how the membrane environment modulates this activity. Therefore, we tested how the yeast plasma membrane P4-ATPase, Dnf2, responds to changes in membrane composition induced by perturbation of endogenous lipid biosynthetic pathways or exogenous application of lipid. The primary substrates of Dnf2 are glucosylceramide (GlcCer) and phosphatidylcholine (PC, or their lyso-lipid derivatives), and we find that these substrates compete with each other for transport. Acutely inhibiting sphingolipid synthesis using myriocin attenuates transport of exogenously applied GlcCer without perturbing PC transport. Deletion of genes controlling later steps of glycosphingolipid production also perturb GlcCer transport to a greater extent than PC transport. In contrast, perturbation of ergosterol biosynthesis reduces PC and GlcCer transport equivalently. Surprisingly, application of lipids that are poor transport substrates differentially affects PC and GlcCer transport by Dnf2, thus altering substrate preference. Our data indicate that Dnf2 exhibits exquisite sensitivity to the membrane composition, thus providing feedback onto the function of the P4-ATPases.

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