The GEF1 Proton-Chloride Exchanger Affects Tombusvirus Replication via Regulation of Copper Metabolism in Yeast

Sasvari, Zsuzsanna; Kovalev, Nikolay; Nagy, Peter D.

doi:10.1128/jvi.02003-12

Cited by 14 publications

(9 citation statements)

References 80 publications

(119 reference statements)

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“…Thus the yeast GEF1 protein is involved in the co-transport of Cu 2+ or Fe 3+ , the regulation of cation homeostasis and the growth of yeast cells. In the Δgef1 loss-of-function yeast mutant, the Cl - transport into the intracellular vesicles (vacuole or Golgi apparatus) was blocked, resulting in hypersensitivity to extracellular cations such as Cu 2+ , Fe 3+ , and Mn 2+ , hygromycin B, tetramethyl ammonium chloride, and thus growth were hindered ( Gaxiola et al, 1998 ; Lv et al, 2009 ; Sasvari et al, 2013 ). Among the seven Arabidopsis CLC paralogs ( AtCLCa-g ), over-expressions of AtCLCa, AtCLCd , and AtCLCf in the yeast gef1 mutant could complement the deficiency in GEF1 protein functions or growth phenotype ( Lv et al, 2009 ; Barbier-Brygo et al, 2011 ).…”

Section: Discussionmentioning

confidence: 99%

GmCLC1 Confers Enhanced Salt Tolerance through Regulating Chloride Accumulation in Soybean

et al. 2016

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The family of chloride channel proteins that mediate Cl- transportation play vital roles in plant nutrient supply, cellular action potential and turgor pressure adjustment, stomatal movement, hormone signal recognition and transduction, Cl- homeostasis, and abiotic and biotic stress tolerance. The anionic toxicity, mainly caused by chloride ions (Cl-), on plants under salt stress remains poorly understood. In this work, we investigated the function of soybean Cl-/H+ antiporter GmCLC1 under salt stress in transgenic Arabidopsis thaliana, soybean, and yeast. We found that GmCLC1 enhanced salt tolerance in transgenic A. thaliana by reducing the Cl- accumulation in shoots and hence released the negative impact of salt stress on plant growth. Overexpression of GmCLC1 in the hairy roots of soybean sequestered more Cl- in their roots and transferred less Cl- to their shoots, leading to lower relative electrolyte leakage values in the roots and leaves. When either the soybean GmCLC1 or the yeast chloride transporter gene, GEF1, was transformed into the yeast gef1 mutant, and then treated with different chloride salts (MnCl2, KCl, NaCl), enhanced survival rate was observed. The result indicates that GmCLC1 and GEF1 exerted similar effects on alleviating the stress of diverse chloride salts on the yeast gef1 mutant. Together, this work suggests a protective function of GmCLC1 under Cl- stress.

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Section: Discussionmentioning

confidence: 99%

GmCLC1 Confers Enhanced Salt Tolerance through Regulating Chloride Accumulation in Soybean

et al. 2016

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“…Follow up experiments using tombusviruses and plants (based on N. benthamiana host) have also showed that many of the identified host factors in yeast are relevant in the plant hosts, too. For example, co-opted host proteins, such as Hsp70, eEF1A, eEF1Bγ, GAPDH, DEAD-box helicases, cellular ion pumps, and ESCRT factors or the roles of sterols and phospholipids have all been confirmed to play important roles in the assembly of the tombusvirus VRCs or viral RNA synthesis in vitro, in yeast and plant cells Jaag et al, 2010;Kovalev and Nagy, 2014;Li et al, 2009;Sasvari et al, 2011;Sasvari et al, 2013;Wang and Nagy, 2008;Wang et al, 2009a;Xu and Nagy, 2015).…”

Section: Discussionmentioning

confidence: 98%

Cellular Ubc2/Rad6 E2 ubiquitin-conjugating enzyme facilitates tombusvirus replication in yeast and plants

et al. 2015

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Mono- and multi-ubiquitination alters the functions and subcellular localization of many cellular and viral proteins. Viruses can co-opt or actively manipulate the ubiquitin network to support viral processes or suppress innate immunity. Using yeast (Saccharomyces cerevisiae) model host, we show that the yeast Rad6p (radiation sensitive 6) E2 ubiquitin-conjugating enzyme and its plant ortholog, AtUbc2, interact with two tombusviral replication proteins and these E2 ubiquitin-conjugating enzymes could be co-purified with the tombusvirus replicase. We demonstrate that TBSV RNA replication and the mono- and bi-ubiquitination level of p33 is decreased in rad6Δ yeast. However, plasmid-based expression of AtUbc2p could complement both defects in rad6Δ yeast. Knockdown of UBC2 expression in plants also decreases tombusvirus accumulation and reduces symptom severity, suggesting that Ubc2p is critical for virus replication in plants. We provide evidence that Rad6p is involved in promoting the subversion of Vps23p and Vps4p ESCRT proteins for viral replicase complex assembly.

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“…By contrast, the ER lumen in the CLC‐Nb1a CLC‐Nb1b mutant could not be modified by PVY infection, resulting in suppression of PVY intracellular replication and systemic movement. In addition to this study, there is some other evidence indicating that CLCs are involved in viral infection (Sasvari et al ., ; Zheng et al ., ; Igloi et al ., ). For example, yeast GEF1 is suggested to affect tombusvirus replication via regulation of copper metabolism (Sasvari et al ., ).…”

Section: Discussionmentioning

confidence: 99%

“…In addition to this study, there is some other evidence indicating that CLCs are involved in viral infection (Sasvari et al ., ; Zheng et al ., ; Igloi et al ., ). For example, yeast GEF1 is suggested to affect tombusvirus replication via regulation of copper metabolism (Sasvari et al ., ). The data presented here support the idea that CLC‐Nt1 affects PVY infection via regulation of ER luminal pH.…”

Section: Discussionmentioning

confidence: 99%

CLC‐Nt1 affects Potato Virus Y infection via regulation of endoplasmic reticulum luminal Ph

et al. 2018

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Chloride channel (CLC) proteins are important anion transporters conserved in organisms ranging from bacteria and yeast to plants and animals. According to sequence comparison, some plant CLCs are predicted to function as Cl /H antiporters, but not Cl channels. However, no direct evidence was provided to verify the role of these plant CLCs in regulating the pH of the intracellular compartment. We identified tobacco CLC-Nt1 interacting with the Potato virus Y (PVY) 6K2 protein. To investigate its physiological function, homologous genes of CLC-Nt1 in Nicotiana benthamiana were knocked out using the CRISPR/Cas9 system. Complementation experiments were subsequently performed by expression of wild-type or point-mutated CLC-Nt1 in knockout mutants. The data presented herein demonstrate that CLC-Nt1 is localized at endoplasmic reticulum (ER). Using a pH-sensitive fluorescent protein (pHluorin), we found that loss of CLC-Nt1 function resulted in a decreased ER luminal pH. Secreted GFP (secGFP) was retained mostly in ER in knockout mutants, indicating that CLC-Nt1 is also involved in protein secretion. PVY infection induced a rise in ER luminal pH, which was dependent on functional CLC-Nt1. By contrast, loss of CLC-Nt1 function inhibited PVY intracellular replication and systemic infection. We propose that PVY alters ER luminal pH for infection in a CLC-Nt1-dependent manner.

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The GEF1 Proton-Chloride Exchanger Affects Tombusvirus Replication via Regulation of Copper Metabolism in Yeast

Cited by 14 publications

References 80 publications

GmCLC1 Confers Enhanced Salt Tolerance through Regulating Chloride Accumulation in Soybean

GmCLC1 Confers Enhanced Salt Tolerance through Regulating Chloride Accumulation in Soybean

Cellular Ubc2/Rad6 E2 ubiquitin-conjugating enzyme facilitates tombusvirus replication in yeast and plants

CLC‐Nt1 affects Potato Virus Y infection via regulation of endoplasmic reticulum luminal Ph

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