scholarly journals TOL proteins mediate vacuolar sorting of the borate transporter BOR1 in Arabidopsis thaliana

2018 ◽  
Author(s):  
Akira Yoshinari ◽  
Barbara Korbei ◽  
Junpei Takano
Keyword(s):  
Arabidopsis Thaliana ◽  
Plasma Membrane ◽  
Intracellular Localization ◽  
The Other ◽  
Dependent Manner ◽  
Root Cells ◽  
Polar Localization ◽  
Other Hand ◽  
Early Endosomes ◽  
Vacuolar Sorting

ABSTRACTBoron (B) is an essential micronutrient for plants, however, it shows cytotoxicity at high concentrations. A borate transporter BOR1 is required for efficient transport of boron (B) toward the root stele in Arabidopsis thaliana. BOR1 shows polar localization in the plasma membrane of various root cells toward the stele-side under B limitation. To avoid over-accumulation of B, BOR1 in the plasma membrane is rapidly internalized and transported into the vacuole for proteolysis after high-B supply in an ubiquitination-dependent manner. Although BOR1 has been predicted to be transported into multi-vesicular bodies/late endosomes (MVB/LEs) via the endosomal sorting complex required for transport (ESCRT) machinery, experimental evidence was absent so far. In this study, we investigated the intracellular localization of BOR1 by visualizing endomembrane compartments, and tested the involvement of ESCRT-0-like proteins TOM1-LIKEs (TOLs) in the vacuolar sorting of BOR1. Under low-B conditions, a large portion of cytoplasmic BOR1 was localized in the trans-Golgi networks/early endosomes (TGN/EEs) labeled with VHA-a1 subunit. Pharmacological interference of endosomal recycling using brefeldin A induced colocalization of BOR1 with RabA5D, which labels recycling vesicles associated with the TGN. These data suggest that BOR1 cycles between plasma membrane and TGN/EE via RabA5D-positive endomembrane compartments under low-B conditions. On the other hand, under high-B conditions, BOR1 was localized in the inside of TOL5-positive MVB/LEs. To examine the roles of TOL proteins in intracellular trafficking of BOR1, we analyzed BOR1-GFP localization in the TOL quintuple mutant (tolQ; tol2-1tol3-1tol5-1tol6-1tol9-1) after high-B supply. In the tolQ mutant, vacuolar sorting of BOR1 was delayed, while the polar localization of BOR1 was not disturbed. Taken together, BOR1 is constantly transported to the TGN/EE by endocytosis and recycled to the plasma membrane likely via RabA5D-positive endomembrane compartments under low-B conditions. On the other hand, BOR1 is transported to the vacuole via TOL5-positive MVB/LEs under high-B conditions. TOL proteins are required for sorting of ubiquitinated BOR1 into MVB/LE for vacuolar degradation but not for the polar trafficking of BOR1.

Translocation of the neuronal cytoskeleton and axonal locomotion

1982 ◽  
Vol 299 (1095) ◽  
pp. 313-327 ◽  
Keyword(s):  
Plasma Membrane ◽  
Axonal Transport ◽  
Cell Body ◽  
Cytoskeletal Proteins ◽  
The Other ◽  
Current Understanding ◽  
Axonal Elongation ◽  
Neuronal Cytoskeleton ◽  
Other Hand ◽  
Neuron Cell Body

Recent studies of axonal transport indicate that cytoskeletal proteins are assembled into polymers in the neuron cell body and that these polymers move from the cell body toward the end of the axon. On the other hand, membranous elements appear to be inserted into the axonal plasma membrane preferentially at the end of the axon. These new observations are explored in relation to our current understanding of axonal elongation.

scholarly journals c-Cbl Mediates Ubiquitination, Degradation, and Down-regulation of Human Protease-activated Receptor 2

2005 ◽  
Vol 280 (16) ◽  
pp. 16076-16087 ◽  
Author(s):  
Claire Jacob ◽  
Graeme S. Cottrell ◽  
Daphne Gehringer ◽  
Fabien Schmidlin ◽  
Eileen F. Grady ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Ring Finger ◽  
Dominant Negative ◽  
Dependent Manner ◽  
Wild Type ◽  
Lysosomal Degradation ◽  
Proteolytic Activation ◽  
Gpcr Signaling ◽  
Early Endosomes ◽  
Protease Activated Receptor 2

Mechanisms that arrest G-protein-coupled receptor (GPCR) signaling prevent uncontrolled stimulation that could cause disease. Although uncoupling from heterotrimeric G-proteins, which transiently arrests signaling, is well described, little is known about the mechanisms that permanently arrest signaling. Here we reported on the mechanisms that terminate signaling by protease-activated receptor 2 (PAR2), which mediated the proinflammatory and nociceptive actions of proteases. Given its irreversible mechanism of proteolytic activation, PAR2is a model to study the permanent arrest of GPCR signaling. By immunoprecipitation and immunoblotting, we observed that activated PAR2was mono-ubiquitinated. Immunofluorescence indicated that activated PAR2translocated from the plasma membrane to early endosomes and lysosomes where it was degraded, as determined by immunoblotting. Mutant PAR2lacking intracellular lysine residues (PAR2Δ14K/R) was expressed at the plasma membrane and signaled normally but was not ubiquitinated. Activated PAR2Δ14K/R internalized but was retained in early endosomes and avoided lysosomal degradation. Activation of wild type PAR2stimulated tyrosine phosphorylation of the ubiquitin-protein isopeptide ligase c-Cbl and promoted its interaction with PAR2at the plasma membrane and in endosomes in an Src-dependent manner. Dominant negative c-Cbl lacking the ring finger domain inhibited PAR2ubiquitination and induced retention in early endosomes, thereby impeding lysosomal degradation. Although wild type PAR2was degraded, and recovery of agonist responses required synthesis of new receptors, lysine mutation and dominant negative c-Cbl impeded receptor ubiquitination and degradation and allowed PAR2to recycle and continue to signal. Thus, c-Cbl mediated ubiquitination and lysosomal degradation of PAR2to irrevocably terminate signaling by this and perhaps other GPCRs.

scholarly journals Endocytic Down-Regulation of ErbB2 Is Stimulated by Cleavage of Its C-Terminus

2007 ◽  
Vol 18 (9) ◽  
pp. 3656-3666 ◽  
Author(s):  
Mads Lerdrup ◽  
Silas Bruun ◽  
Michael V. Grandal ◽  
Kirstine Roepstorff ◽  
Malene M. Kristensen ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Retention Mechanism ◽  
Full Length ◽  
Dependent Manner ◽  
Lysosomal Degradation ◽  
Down Regulation ◽  
C Terminus ◽  
Early Endosomes

High ErbB2 levels are associated with cancer, and impaired endocytosis of ErbB2 could contribute to its overexpression. Therefore, knowledge about the mechanisms underlying endocytic down-regulation of ErbB2 is warranted. The C-terminus of ErbB2 can be cleaved after various stimuli, and after inhibition of HSP90 with geldanamycin this cleavage is accompanied by proteasome-dependent endocytosis of ErbB2. However, it is unknown whether C-terminal cleavage is linked to endocytosis. To study ErbB2 cleavage and endocytic trafficking, we fused yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP) to the N- and C-terminus of ErbB2, respectively (YFP-ErbB2-CFP). After geldanamycin stimulation YFP-ErbB2-CFP became cleaved in nonapoptotic cells in a proteasome-dependent manner, and a markedly larger relative amount of cleaved YFP-ErbB2-CFP was observed in early endosomes than in the plasma membrane. Furthermore, cleavage took place at the plasma membrane, and cleaved ErbB2 was internalized and degraded far more efficiently than full-length ErbB2. Concordantly, a C-terminally truncated ErbB2 was also readily endocytosed and degraded in lysosomes compared with full-length ErbB2. Altogether, we suggest that geldanamycin leads to C-terminal cleavage of ErbB2, which releases the receptor from a retention mechanism and causes endocytosis and lysosomal degradation of ErbB2.

scholarly journals Antiausterity Activity of Arctigenin Enantiomers: Importance of (2R,3R)-Absolute Configuration

2014 ◽  
Vol 9 (1) ◽  
pp. 1934578X1400900 ◽  
Author(s):  
Suresh Awale ◽  
Mamoru Kato ◽  
Dya Fita Dibwe ◽  
Feng Li ◽  
Chika Miyoshi ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Cancer Cells ◽  
Absolute Configuration ◽  
The Other ◽  
Dependent Manner ◽  
Meoh Extract ◽  
Concentration Dependent Manner ◽  
Akt Activation ◽  
Other Hand

From a MeOH extract of powdered roots of Wikstroema indica, six dibenzyl-γ-butyrolactone-type lignans with (2 S,3 S)-absolute configuration [(+)-arctigenin (1), (+)-matairesinol (2), (+)-trachelogenin (3), (+)-nortrachelogenin (4), (+)-hinokinin (5), and (+)-kusunokinin (6)] were isolated, whereas three dibenzyl-γ-butyrolactone-type lignans with (2 R,3 R)-absolute configuration [(-)-arctigenin (1), (-)-matairesinol (2), (-)-trachelogenin (3)] were isolated from Trachelospermum asiaticum. The in vitro preferential cytotoxic activity of the nine compounds was evaluated against human pancreatic PANC-1 cancer cells in nutrient-deprived medium (NDM), but none of the six lignans (1–6) with (2 S,3 S)-absolute configuration showed preferential cytotoxicity. On the other hand, three lignans (1*–3*) with (2 R,3 R)-absolute configuration exhibited preferential cytotoxicity in a concentration-dependent manner with PC50 values of 0.54, 6.82, and 5.85 μM, respectively. Furthermore, the effect of (-)- and (+)-arctigenin was evaluated against the activation of Akt, which is a key process in the tolerance to nutrition starvation. Interestingly, only (-)-arctigenin (1*) strongly suppressed the activation of Akt. These results indicate that the (2 R,3 R)-absolute configuration of (-)-enantiomers should be required for the preferential cytotoxicity through the inhibition of Akt activation.

Effects of Trehalose and Ethanol on Yeast Cytosolic Pyrophosphatase

1999 ◽  
Vol 54 (3-4) ◽  
pp. 186-190 ◽  
Author(s):  
Dahabada Helena José Lopes ◽  
José Roberto Meyer-Fernandes ◽  
Mauro Sola-Penna
Keyword(s):  
Fermentation Process ◽  
Stress Condition ◽  
The Other ◽  
Dependent Manner ◽  
Control Activity ◽  
Other Hand ◽  
Ethanol Ethanol ◽  
Stress Protectant ◽  
Dose Dependent

Trehalose has been described to protect several enzymes against destabilizing conditions. This sugar is naturally accumulated by yeast as a stress protectant. A common stress condition that yeast is normally submitted is the presence of ethanol, the by-product of fermentation process of several yeast. In this paper we show the effects of trehalose and ethanol, alone or together, on yeast pyrophosphatase, and the effects of these compounds on inhibition and unfolding of pyrophosphatase promoted by urea. We show that both trehalose and ethanol inhibit pyrophosphatase in a dose-dependent manner, and that the presence of ethanol does not modify the inhibition promoted by trehalose as well as the presence of trehalose does not modify the inhibition promoted by ethanol. The effects of trehalose on pyrophosphatase are completely reversible, but the inhibition caused by ethanol was only partially reversible. Incubation of pyrophosphatase with 10% (v/v) ethanol promoted an inhibition of 15%, and the control activity was completely recovered after removal of ethanol. On the other hand, when pyrophosphatase was incubated with 20% (v/v) ethanol an inhibition of 40% of the control activity was observed which persisted after removal of ethanol. Ethanol also potentiates the inhibition of pyrophosphatase promoted by urea, and contributes for an irreversible inactivation and unfolding of pyrophosphatase in the presence of urea. Trehalose, that protects this enzyme against the inhibition and unfolding promoted by the chaotropic compound urea, was inefficient to protect against the effects of ethanol. Trehalose was also efficient to prevent an irreversible inactivation induced by urea.

scholarly journals The BLOC-1 complex promotes endosomal maturation by recruiting the Rab5 GTPase-activating protein Msb3

2013 ◽  
Vol 201 (1) ◽  
pp. 97-111 ◽  
Author(s):  
Arun T. John Peter ◽  
Jens Lachmann ◽  
Meenakshi Rana ◽  
Madeleine Bunge ◽  
Margarita Cabrera ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Protein Sorting ◽  
Early Endosome ◽  
Endocytic Pathway ◽  
Dependent Manner ◽  
Gtpase Activating Protein ◽  
Early Endosomes ◽  
Lysosome Related Organelles

Membrane microcompartments of the early endosomes serve as a sorting and signaling platform, where receptors are either recycled back to the plasma membrane or forwarded to the lysosome for destruction. In metazoan cells, three complexes, termed BLOC-1 to -3, mediate protein sorting from the early endosome to lysosomes and lysosome-related organelles. We now demonstrate that BLOC-1 is an endosomal Rab-GAP (GTPase-activating protein) adapter complex in yeast. The yeast BLOC-1 consisted of six subunits, which localized interdependently to the endosomes in a Rab5/Vps21-dependent manner. In the absence of BLOC-1 subunits, the balance between recycling and degradation of selected cargoes was impaired. Additionally, our data show that BLOC-1 is both a Vps21 effector and an adapter for its GAP Msb3. BLOC-1 and Msb3 interacted in vivo, and both mutants resulted in a redistribution of active Vps21 to the vacuole surface. We thus conclude that BLOC-1 controls the lifetime of active Rab5/Vps21 and thus endosomal maturation along the endocytic pathway.

Functional Pool of Cyclic Adenosine 3',5'-Monophosphate in Rabbit Platelets

1983 ◽  
Vol 49 (01) ◽  
pp. 008-012 ◽  
Author(s):  
Shuichi G Hashimoto
Keyword(s):  
Plasma Membrane ◽  
Cyclic Amp ◽  
Binding Capacity ◽  
Binding Activity ◽  
The Other ◽  
Rabbit Platelet ◽  
Platelet Membranes ◽  
Other Hand ◽  
Fraction I ◽  
Induced Aggregation

SummaryAccumulation of the newly formed 14C-cyclic adenosine 3',5’-monophosphate (cyclic AMP) was found in the P1 (1.0) fraction, i. e. a platelet plasma membrane fraction which was obtained from 14C-adenine-labeled platelets. On the other hand, total cyclic AMP as determined simultaneously was located mainly in the platelet soluble fraction. Furthermore, the highest value of the cyclic AMP-binding capacity was found in the P1 (1.0) fraction. The cyclic AMP-binding activity of platelet membranes was attributed to two proteins with molecular weights of approximately 48,000 and 68,000.The treatment of 14C-adenine-prelabeled platelets with thrombin (1 unit per ml) led to about 40% decrease in the newly formed 14C-cyclic AMP level and 18% reduction of 14C-adenosine triphosphate level in whole platelets within 10 sec. On the other hand, the 14C-cyclic AMP level in the P, fraction decreased by about 80% of the control value while the total cyclic AMP in this fraction was almost unchanged. This rapid and striking fall in the membrane 14C-cyclic AMP level could be correlated with the more than 2fold stimulation of the membrane-bound cyclic AMP phosphodiesterase, together with the more than 20% inhibition of both the cyclic AMP-binding capacity and the adenyl cyclase in platelet membranes by thrombin treatment. These observations suggest the possibility that functional pool of cyclic AMP related to thrombin-induced aggregation is located in rabbit platelet plasma membrane.

scholarly journals Suppression of progesterone-enhanced hyperactivation in hamster spermatozoa by estrogen

Reproduction ◽  
2010 ◽  
Vol 140 (3) ◽  
pp. 453-464 ◽  
Author(s):  
Masakatsu Fujinoki
Keyword(s):  
Steroid Hormones ◽  
Sperm Head ◽  
The Other ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Other Hand ◽  
Sperm Proteins ◽  
17Β Estradiol

In this study, I examined whether sperm hyperactivation in hamster is regulated by steroid hormones such as estrogen (estradiol, E2) and progesterone. Although sperm hyperactivation was enhanced by progesterone, 17β-estradiol (17βE2) itself did not affect sperm hyperactivation. However, 17βE2 suppressed progesterone-enhanced hyperactivation in a concentration-dependent manner through non-genomic pathways when spermatozoa were exposed to 17βE2 at the same time or before exposure to progesterone. When spermatozoa were exposed to 17βE2 after exposure to progesterone, 17βE2 did not suppress progesterone-enhanced hyperactivation. Moreover, 17α-estradiol, an inactive isomer of E2, did not suppress progesterone-enhanced hyperactivation. Observations using a FITC-conjugated 17βE2 showed that it binds to the acrosome region of the sperm head. Binding of 17βE2 to spermatozoa was not inhibited by progesterone, although 17βE2 did not suppress progesterone-enhanced hyperactivation when spermatozoa were exposed to 17βE2 after exposure to progesterone. On the other hand, binding of progesterone to spermatozoa was also not inhibited by 17βE2 even if progesterone-enhanced hyperactivation was suppressed by 17βE2. Although tyrosine phosphorylations of sperm proteins were enhanced by progesterone, enhancement of tyrosine phosphorylations by progesterone was suppressed by 17βE2. Moreover, tyrosine phosphorylations were inhibited by 17βE2 when only 17βE2 was added to the medium. From these results, it is likely that 17βE2 competitively suppresses progesterone-enhanced hyperactivation through the inhibition of tyrosine phosphorylations via non-genomic pathways.

Serotonin relaxes porcine pial veins

1994 ◽  
Vol 266 (3) ◽  
pp. H1000-H1006 ◽  
Author(s):  
T. J. Lee ◽  
M. Ueno ◽  
N. Sunagane ◽  
M. H. Sun
Keyword(s):  
Muscle Tone ◽  
The Other ◽  
Receptor Subtypes ◽  
Dependent Manner ◽  
Pial Arteries ◽  
Concentration Dependent Manner ◽  
Vasoactive Substances ◽  
Other Hand ◽  
Mechanical Stretching ◽  
Or Applications

The effect of serotonin [5-hydroxytryptamine (5-HT)] on pial venous tone of the pig was examined using in vitro tissue bath techniques. Isolated pial venous rings exhibited spontaneous rhythmic contractions (SRC) on mechanical stretching and/or applications of several vasoactive substances, including norepinephrine. On the other hand, KCl induced sustained active muscle tone (SAT) without SRC. The SRC induced by mechanical stretching were not affected by tetrodotoxin, nitro-L-arginine, alpha- and beta-adrenergic, histaminergic, and muscarinic receptor antagonists, indicating that the SRC in porcine pial veins are of myogenic origin. The SRC induced by stretching or applications of vasoactive substances and SAT induced by KCl were inhibited by 5-HT in a concentration-dependent manner. The inhibition was prevented by methysergide and methiothepin but not by ketanserin, propranolol, 3 alpha-tropanyl-1H-indole-3-carboxylic acid ester, hemoglobin, or nitro-L-arginine. The SRC and SAT were inhibited by 5-carboxamidotryptamine (5-CT), 8-hydroxy-2-di-N-propylaminotetralin HBr (8-OHDPAT), 1-[3-(trifluoromethyl)phenyl]piperazine (TFMPP), and 5-methoxytryptamine (5-MT), but not by sumatriptan, alpha-methylserotonin, or 2-methylserotonin. On the other hand, 5-CT, 8-OHDPAT, TFMPP, 5-MT, and sumatriptan constricted the porcine pial arteries exclusively. In 15% of pial venous preparations examined, 5-HT at low concentrations induced ketanserin-sensitive constrictions. These results indicate that the porcine pial venous smooth muscle contains multiple subtypes of 5-HT receptors. The 5-HT inhibition of SRC and SAT is predominant and is mediated by 5-HT1-like receptors, which, however, do not seem to correspond to 5-HT1A, 5-HT1B, 5-HT1C, 5-HT1D, 5-HT1E, or 5-HT1F receptor subtypes.(ABSTRACT TRUNCATED AT 250 WORDS)

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