共查询到20条相似文献,搜索用时 15 毫秒
1.
Ernst S Zobiack N Boecker K Gerke V Rescher U 《Cellular and molecular life sciences : CMLS》2004,61(13):1684-1692
The formyl peptide-like receptor FPRL1 is a member of the chemoattractant subfamily of G protein- coupled receptors involved in regulating leukocyte migration in inflammation. To elucidate mechanisms underlying the internalization of ligand-bound FPRL1 and possible receptor recycling, we characterized the endocytic itinerary of FPRL1. We show that agonist-triggered internalization from the plasma membrane into intracellular compartments is prevented by perturbation of clathrin-mediated endocytosis, such as expression of the dominant-negative clathrin Hub mutant, siRNA-mediated depletion of cellular clathrin and expression of a dominant-negative mutant of the large GTPase dynamin. Internalized FPRL1 co-localized with endocytosed transferrin and the small GTPases Rab4 and Rab11 in vesicular structures most resembling recycling endosomes. Recycling of FPRL1 was significantly reduced by pretreatment with PI3-kinase inhibitors. Thus, ligand-bound FPRL1 undergoes primarily clathrin-mediated and dynamin-dependent endocytosis and the receptor recycles via a rapid PI3-kinase-sensitive route as well as pathways involving perinuclear recycling endosomes.Received 19 March 2004; received after revision 26 April 2004; accepted 12 May 2004 相似文献
2.
Iness Charfi Khaled Abdallah Louis Gendron Graciela Pineyro 《Cellular and molecular life sciences : CMLS》2018,75(12):2257-2271
Soon after internalization delta opioid receptors (DOPrs) are committed to the degradation path by G protein-coupled receptor (GPCR)-associated binding protein. Here we provide evidence that this classical post-endocytic itinerary may be rectified by downstream sorting decisions which allow DOPrs to regain to the membrane after having reached late endosomes (LE). The LE sorting mechanism involved ESCRT accessory protein Alix and the TIP47/Rab9 retrieval complex which supported translocation of the receptor to the TGN, from where it subsequently regained the cell membrane. Preventing DOPrs from completing this itinerary precipitated acute analgesic tolerance to the agonist DPDPE, supporting the relevance of this recycling path in maintaining the analgesic response by this receptor. Taken together, these findings reveal a post-endocytic itinerary where GPCRs that have been sorted for degradation can still recycle to the membrane. 相似文献
3.
Alvi F Idkowiak-Baldys J Baldys A Raymond JR Hannun YA 《Cellular and molecular life sciences : CMLS》2007,64(3):263-270
The protein kinase C (PKC) family of isoenzymes has been shown to regulate a variety of cellular processes, including receptor
desensitization and internalization, and this has sparked interest in further delineation of the roles of specific isoforms
of PKC in membrane trafficking and endocytosis. Recent studies have identified a novel translocation of PKC to a juxtanuclear
compartment, the pericentrion, which is distinct from the Golgi complex but epicentered on the centrosome. Sustained activation
of PKC (longer than 30 min) also results in sequestration of plasma membrane lipids and proteins to the same compartment,
demonstrating a global effect on endocytic trafficking. This review summarizes these studies, particularly focusing on the
characterization of the pericentrion as a distinct PKC-dependent subset of recycling endosomes. We also discuss emerging insights
into a role for PKC as a central hub in regulating vesicular transport pathways throughout the cell, with implications for
a wide range of pathobiologic processes, e.g. diabetes and abnormal neurotransmission or receptor desensitization.
Received 11 August 2006; received after revision 20 September 2006; accepted 7 November 2006 相似文献
4.
Uusi-Rauva K Kyttälä A van der Kant R Vesa J Tanhuanpää K Neefjes J Olkkonen VM Jalanko A 《Cellular and molecular life sciences : CMLS》2012,69(12):2075-2089
CLN3 is an endosomal/lysosomal transmembrane protein mutated in classical juvenile onset neuronal ceroid lipofuscinosis, a fatal inherited neurodegenerative lysosomal storage disorder. The function of CLN3 in endosomal/lysosomal events has remained elusive due to poor understanding of its interactions in these compartments. It has previously been shown that the localisation of late endosomal/lysosomal compartments is disturbed in cells expressing the most common disease-associated CLN3 mutant, CLN3?ex7-8 (c.462-677del). We report here that a protracted disease causing mutant, CLN3E295K, affects the properties of late endocytic compartments, since over-expression of the CLN3E295K mutant protein in HeLa cells induced relocalisation of Rab7 and a perinuclear clustering of late endosomes/lysosomes. In addition to the previously reported disturbances in the endocytic pathway, we now show that the anterograde transport of late endosomal/lysosomal compartments is affected in CLN3 deficiency. CLN3 interacted with motor components driving both plus and minus end microtubular trafficking: tubulin, dynactin, dynein and kinesin-2. Most importantly, CLN3 was found to interact directly with active, guanosine-5'-triphosphate (GTP)-bound Rab7 and with the Rab7-interacting lysosomal protein (RILP) that anchors the dynein motor. The data presented in this study provide novel insights into the role of CLN3 in late endosomal/lysosomal membrane transport. 相似文献
5.
Macroautophagy, the process by which cytosolic components and organelles are engulfed and degraded by a double-membrane structure,
could be viewed as a specialized, multistep membrane transport process. As such, it intersects with the exocytic and endocytic
membrane trafficking pathways. A number of Rab GTPases which regulate secretory and endocytic membrane traffic have been shown
to play either critical or accessory roles in autophagy. The biogenesis of the pre-autophagosomal isolation membrane (or phagophore)
is dependent on the functionality of Rab1. A non-canonical, Atg5/Atg7-independent mode of autophagosome generation from the
trans-Golgi or endosome requires Rab9. Other Rabs, such as Rab5, Rab24, Rab33, and Rab7 have all been shown to be required,
or involved at various stages of autophagosomal genesis and maturation. Another small GTPase, RalB, was very recently demonstrated
to induce isolation membrane formation and maturation via its engagement of the exocyst complex, a known Rab effector. We
summarize here what is now known about the involvement of Rabs in autophagy, and discuss plausible mechanisms with future
perspectives. 相似文献
6.
Tulapurkar ME Schäfer R Hanck T Flores RV Weisman GA González FA Reiser G 《Cellular and molecular life sciences : CMLS》2005,62(12):1388-1399
Extracellular nucleotides exert a large number of physiological effects through activation of P2Y receptors. We expressed rat P2Y2 (rP2Y2) receptor, tagged with green fluorescent protein (GFP) in HEK-293 cells and visualized receptor translocation in live cells by confocal microscopy. Functional receptor expression was confirmed by determining [Ca2+]i responses. Agonist stimulation caused a time-dependent translocation of the receptor from the plasma membrane to the cytoplasm. Rearrangement of the actin cytoskeleton was observed during agonist-mediated rP2Y2-GFP receptor internalization. Colocalization of the internalized receptor with early endosomes, clathrin and lysosomes was detected by confocal microscopy. The inhibition of receptor endocytosis by either high-density medium or chlorpromazine in the presence of UTP indicates that the receptor was internalized by the clathrin-mediated pathway. The caveolin- mediated pathway was not involved. Targeting of the receptor from endosomes to lysosomes seems to involve the proteasome pathway, because proteasomal inhibition increased receptor recycling back to the plasma membrane.Received 8 February 2005; received after revision 18 March 2005; accepted 11 April 2005 相似文献
7.
Spampinato S Di Toro R Alessandri M Murari G 《Cellular and molecular life sciences : CMLS》2002,59(12):2172-2183
In this study, we examined agonist-induced internalization, recycling and signalling (measure of cAMP levels) of the cloned human nociceptin receptor (hNOP) expressed in CHO-K1 cells. Internalization was proven by a receptor-binding assay on viable cells. The agonist nociceptin/orphanin FQ (NC) promoted rapid internalization of the hNOP receptor (approximately 78% of cell surface receptors were lost after 2 min exposure to 1 microM NC) in a clathrin- and ATP-dependent manner. Internalization was more rapid and marked in CHO-K1 cells than, as we previously reported, in SK-N-BE cells. This difference may be related to higher levels of beta-arrestin isoforms detected in CHO-K1 than in SK-N-BE cells. hNOP receptor internalization was partially reversible and recycling occurred in the presence of the agonist; receptor recycling was dependent on okadaic acid-sensitive phosphatases and was blocked by monensin. Confocal microscopy analysis confirmed the internalization and the recycling back to the plasma membrane of an epitope-tagged hNOP receptor expressed in CHO-K1 cells. These receptors underwent rapid desensitization upon agonist challenge: NC efficacy in inhibiting forskolin-stimulated cAMP production was significantly reduced 10 min after exposure and correlated with the rate of receptor internalization. Moreover, we observed that blockade of hNOP receptor recycling by monensin would cause a more prolonged and relevant desensitization of this receptor. Thus, the dynamic cycle between hNOP receptor activation, internalization and recycling determines the activity of this receptor on the cell surface. 相似文献
8.
Sabine Gilch Christian Bach Gloria Lutzny Ina Vorberg Hermann M. Schätzl 《Cellular and molecular life sciences : CMLS》2009,66(24):3979-3991
The infectious agent in prion diseases consists of an aberrantly folded isoform of the cellular prion protein (PrPc), termed PrPSc, which accumulates in brains of affected individuals. Studies on prion-infected cultured cells indicate that cellular cholesterol
homeostasis influences PrPSc propagation. Here, we demonstrate that the cellular PrPSc content decreases upon accumulation of cholesterol in late endosomes, as induced by NPC-1 knock-down or treatment with U18666A.
PrPc trafficking, lipid raft association, and membrane turnover are not significantly altered by such treatments. Cellular PrPSc formation is not impaired, suggesting that PrPSc degradation is increased by intracellular cholesterol accumulation. Interestingly, PrPSc propagation in U18666A-treated cells was partially restored by overexpression of rab 9, which causes redistribution of cholesterol
and possibly of PrPSc to the trans-Golgi network. Surprisingly, rab 9 overexpression itself reduced cellular PrPSc content, indicating that PrPSc production is highly sensitive to alterations in dynamics of vesicle trafficking. 相似文献
9.
Mari SA Soragna A Castagna M Santacroce M Perego C Bossi E Peres A Sacchi VF 《Cellular and molecular life sciences : CMLS》2006,63(1):100-111
We investigated the role of the Q291 glutamine residue in the functioning of the rat γ-aminobutyric acid (GABA) transporter
GAT-1. Q291 mutants cannot transport GABA or give rise to transient, leak and transport-coupled currents even though they
are targeted to the plasma membrane. Coexpression experiments of wild-type and Q291 mutants suggest that GAT-1 is a functional
monomer though it requires oligomeric assembly for membrane insertion. We determined the accessibility of Q291 by investigating
the impact of impermeant sulfhydryl reagents on cysteine residues engineered in close proximity to Q291. The effect of these
reagents indicates that Q291 faces the external aqueous milieu. The introduction of a steric hindrance close to Q291 by means
of [2-(trimethylammonium)ethyl] methanethiosulfonate bromide modification of C74A/T290C altered the affinity of the mutant
for cations. Taken together, these results suggest that this irreplaceable residue is involved in the interaction with sodium
or in maintaining the cation accessibility to the transporter.
Received 24 October 2005; accepted 11 November 2005 相似文献
10.
Lukasz Wujak Ralph T. Böttcher Oleg Pak Helena Frey Elie El Agha Ying Chen Sigrid Schmitt Saverio Bellusci Liliana Schaefer Norbert Weissmann Reinhard Fässler Malgorzata Wygrecka 《Cellular and molecular life sciences : CMLS》2018,75(9):1671-1685
Low density lipoprotein receptor-related protein (LRP) 1 modulates cell adhesion and motility under normal and pathological conditions. Previous studies documented that LRP1 binds several integrin receptors and mediates their trafficking to the cell surface and endocytosis. However, the mechanism by which LRP1 may regulate integrin activation remains unknown. Here we report that LRP1 promotes the activation and subsequent degradation of β1 integrin and thus supports cell adhesion, spreading, migration and integrin signaling on fibronectin. LRP1 interacts with surface β1 integrin, binds the integrin activator kindlin2 and stimulates β1 integrin–kindlin2 complex formation. Specifically, serine 76 in the LRP1 cytoplasmic tail is crucial for the interaction with kindlin2, β1 integrin activation and cell adhesion. Interestingly, a loss of LRP1 induces the accumulation of several integrin receptors on the cell surface. Following internalization, intracellular trafficking of integrins is driven by LRP1 in a protein kinase C- and class II myosin-dependent manner. Ultimately, LRP1 dictates the fate of endocytosed β1 integrin by directing it down the pathway of lysosomal and proteasomal degradation. We propose that LRP1 mediates cell adhesion by orchestrating a multi-protein pathway to activate, traffic and degrade integrins. Thus, LRP1 may serve as a focal point in the integrin quality control system to ensure a firm connection to the extracellular matrix. 相似文献
11.
Konstantina Katsarou Alexandros Α. Lavdas Panagiota Tsitoura Elisavet Serti Panagiotis Markoulatos Penelope Mavromara Urania Georgopoulou 《Cellular and molecular life sciences : CMLS》2010,67(14):2491-2506
Although HCV is an enveloped virus, naked nucleocapsids have been reported in the serum of infected patients. The HCV core
particle serves as a protective capsid shell for the viral genome and recombinant in vitro assembled HCV core particles induce
strong specific immunity. We investigated the post-binding mechanism of recombinant core particle uptake and its intracellular
fate. In hepatic cells, these particles are internalized, most likely in a clathrin-dependent pathway, reaching early to late
endosomes and finally lysosomes. The endocytic acidic milieu is implicated in trafficking process. Using specific phosphoantibodies,
signaling pathway inhibitors and chemical agents, ERK1/2 was found to be activated in a sustained way after endocytosis, followed by downstream immediate early genes (c-fos and egr-1) modulation. We propose that the intriguing properties of cellular internalization of HCV non-enveloped particles can induce
specific ERK1/2–MAPKs events that could be important in HCV life cycle and pathogenesis of HCV infection. 相似文献
12.
Julia Scharnert Lilo Greune Dagmar Zeuschner Marie-Luise Lubos M. Alexander Schmidt Christian Rüter 《Cellular and molecular life sciences : CMLS》2013,70(24):4809-4823
Extracellular Gram-negative pathogenic bacteria target essential cytoplasmic processes of eukaryotic cells by using effector protein delivery systems such as the type III secretion system (T3SS). These secretion systems directly inject effector proteins into the host cell cytoplasm. Among the T3SS-dependent Yop proteins of pathogenic Yersinia, the function of the effector protein YopM remains enigmatic. In a recent study, we demonstrated that recombinant YopM from Yersinia enterocolitica enters host cells autonomously without the presence of bacteria and thus identified YopM as a novel bacterial cell-penetrating protein. Following entry YopM down-regulates expression of pro-inflammatory cytokines such as tumor necrosis factor α. These properties earmark YopM for further development as a novel anti-inflammatory therapeutic. To elucidate the uptake and intracellular targeting mechanisms of this bacterial cell-penetrating protein, we analyzed possible routes of internalization employing ultra-cryo electron microscopy. Our results reveal that under physiological conditions, YopM enters cells predominantly by exploiting endocytic pathways. Interestingly, YopM was detected free in the cytosol and inside the nucleus. We could not observe any colocalization of YopM with secretory membranes, which excludes retrograde transport as the mechanism for cytosolic release. However, our findings indicate that direct membrane penetration and/or an endosomal escape of YopM contribute to the cytosolic and nuclear localization of the protein. Surprisingly, even when endocytosis is blocked, YopM was found to be associated with endosomes. This suggests an intracellular endosome-associated transport of YopM. 相似文献
13.
Aamir S. Mukadam Sophia Y. Breusegem Matthew N. J. Seaman 《Cellular and molecular life sciences : CMLS》2018,75(14):2613-2625
The processing of amyloid precursor protein (APP) to the neurotoxic pro-aggregatory Aβ peptide is controlled by the mechanisms that govern the trafficking and localisation of APP. We hypothesised that genes involved in endosomal protein sorting could play an important role in regulating APP processing and, therefore, analysed ~ 40 novel endosome-to-Golgi retrieval genes previously identified in a genome-wide siRNA screen. We report that phospholipase D3 (PLD3), a type II membrane protein, functions in endosomal protein sorting and plays an important role in regulating APP processing. PLD3 co-localises with APP in endosomes and loss of PLD3 function results in reduced endosomal tubules, impaired trafficking of several membrane proteins and reduced association of sortilin-like 1 with APP. 相似文献
14.
Helin Räägel Margot Hein Asko Kriiska Pille Säälik Anders Florén Ülo Langel Margus Pooga 《Cellular and molecular life sciences : CMLS》2013,70(24):4825-4839
Since their discovery, cell-penetrating peptides (CPPs) have provided a novel, efficient, and non-invasive mode of transport for various (bioactive) cargos into cells. Despite the ever-growing number of successful implications of the CPP-mediated delivery, issues concerning their intracellular trafficking, significant targeting to degradative organelles, and limited endosomal escape are still hindering their widespread use. To overcome these obstacles, we have utilized a potent photo-induction technique with a fluorescently labeled protein cargo attached to an efficient CPP, TP10. In this study we have determined some key requirements behind this induced escape (e.g., dependence on peptide-to-cargo ratio, time and cargo), and have semi-quantitatively assessed the characteristics of the endosomes that become leaky upon this treatment. Furthermore, we provide evidence that the photo-released cargo remains intact and functional. Altogether, we can conclude that the photo-induced endosomes are specific large complexes-condensed non-acidic vesicles, where the released cargo remains in its native intact form. The latter was confirmed with tubulin as the cargo, which upon photo-induction was incorporated into microtubules. Because of this, we propose that combining the CPP-mediated delivery with photo-activation technique could provide a simple method for overcoming major limitations faced today and serve as a basis for enhanced delivery efficiency and a subsequent elevated cellular response of different bioactive cargo molecules. 相似文献
15.
CFTR biogenesis starts with its co-translational insertion into the membrane of endoplasmic reticulum and folding of the cytosolic domains, towards the acquisition of a fully folded compact native structure. Efficiency of this process is assessed by the ER quality control system that allows the exit of folded proteins but targets unfolded/misfolded CFTR to degradation. If allowed to leave the ER, CFTR is modified at the Golgi and reaches the post-Golgi compartments to be delivered to the plasma membrane where it functions as a cAMP- and phosphorylation-regulated chloride/bicarbonate channel. CFTR residence at the membrane is a balance of membrane delivery, endocytosis, and recycling. Several adaptors, motor, and scaffold proteins contribute to the regulation of CFTR stability and are involved in continuously assessing its structure through peripheral quality control systems. Regulation of CFTR biogenesis and traffic (and its dysregulation by mutations, such as the most common F508del) determine its overall activity and thus contribute to the fine modulation of chloride secretion and hydration of epithelial surfaces. This review covers old and recent knowledge on CFTR folding and trafficking from its synthesis to the regulation of its stability at the plasma membrane and highlights how several of these steps can be modulated to promote the rescue of mutant CFTR. 相似文献
16.
Pan Q Qiao F Gao C Norman B Optican L Zelenka PS 《Cellular and molecular life sciences : CMLS》2011,68(20):3425-3436
The non-receptor tyrosine kinase Src is a critical regulator of cytoskeletal contraction, cell adhesion, and migration. In
normal cells, Src activity is stringently controlled by Csk-dependent phosphorylation of Src(Y530), and by Cullin-5-dependent
ubiquitinylation, which affects active Src(pY419) exclusively, leading to its degradation by the proteosome. Previous work
has shown that Src activity is also limited by Cdk5, a proline-directed kinase, which has been shown to phosphorylate Src(S75).
Here we show that this phosphorylation promotes the ubiquitin-dependent degradation of Src, thus restricting the availability
of active Src. We demonstrate that Src(S75) phosphorylation occurs in vivo in epithelial cells, and like ubiquitinylation,
is associated only with active Src. Preventing Cdk5-dependent phosphorylation of Src(S75), by site-specific mutation of S75
or by Cdk5 inhibition or suppression, increases Src(Y419) phosphorylation and kinase activity, resulting in Src-dependent
cytoskeletal changes. In transfected cells, ubiquitinylation of Src(S75A) is about 35% that of wild-type Src-V5, and its half-life
is approximately 2.5-fold greater. Cdk5 suppression leads to a comparable decrease in the ubiquitinylation of endogenous Src
and a similar increase in Src stability. Together, these findings demonstrate that Cdk5-dependent phosphorylation of Src(S75)
is a physiologically significant mechanism of regulating intracellular Src activity. 相似文献
17.
Modification of ligand-gated receptor function at the postsynaptic domain is one of the most important mechanisms by which
the efficacy of synaptic transmission in the nervous system is regulated. Traditionally, these types of modifications have
been thought to be achieved mainly by altering the channel-gating properties or conductance of the receptors. However, recent
evidence suggests that AMPA (α-amino-3-hydroxyl-5-methyl-4-isoxayolepropionic acid)-type ligand-gated glutamate receptors are continuously recycling between
the plasma membrane and the intracellular compartments via vesicle-mediated plasma membrane insertion and clathrin-dependent
endocytosis. Regulation of either receptor insertion or endocytosis results in a rapid change in the number of these receptors
expressed on the plasma membrane surface and in the receptor-mediated responses, thereby playing an important role in mediating
certain forms of synaptic plasticity. Thus, controlling the number of postsynaptic receptors by regulating the intracellular
trafficking and plasma membrane expression of the postsynaptic receptors may be a common and important mechanism of synaptic
plasticity in the mammalian central nervous system. 相似文献
18.
Madeleine Lamborot A. Espinoza E. Alvarez 《Cellular and molecular life sciences : CMLS》1979,35(5):593-595
Summary Most of 12 taxa karyotypes retain 6 pairs of metacentric macrochromosomes (primitive), but show reduced numbers of microchromosomes (2n=34, 32 and 30). Others whow increased diploid numbers due to macrochromosomal fissions (up to 4 fissions, 2n=40). One shows a fission polymorphism.We thank Dr. W.P. Hall for his many suggestions and comments; and L. Alvarez, E. Barrientos, I. Campos, M. Fernandez and F.N. Manzur for generous assistance. This study was aided by the Project del Servicio de Desarrollo Científico de la Universidad de Chile, No. B259-783. 相似文献
19.
Chiow KH Tan Y Chua RY Huang D Ng ML Torta F Wenk MR Wong SH 《Cellular and molecular life sciences : CMLS》2012,69(9):1505-1521
Since being introduced globally as aspirin in 1899, acetylsalicylic acid has been widely used as an analgesic, anti-inflammation,
anti-pyretic, and anti-thrombotic drug for years. Aspirin had been reported to down-regulate surface expression of CD40, CD80,
CD86, and MHCII in myeloid dendritic cells (DC), which played essential roles in regulating the immune system. We hypothesized
that the down-regulation of these surface membrane proteins is partly due to the ability of aspirin in regulating trafficking/sorting
of endocytosed surface membrane proteins. By using an established epidermoid carcinoma cell line (A-431), which overexpresses
the epidermal growth factor receptor (EGFR) and transferrin receptor (TfnR), we show that aspirin (1) reduces cell surface
expression of EGFR and (2) accumulates endocytosed-EGFR and -TfnR in the early/sorting endosome (ESE). Further elucidation
of the mechanism suggests that aspirin enhances recruitment of SNX3 and SNX5 to membranes and consistently, both SNX3 and
SNX5 play essential roles in the aspirin-mediated accumulation of endocytosed-TfnR at the ESE. This study sheds light on how
aspirin may down-regulate surface expression of EGFR by inhibiting/delaying the exit of endocytosed-EGFR from the ESE and
recycling of endocytosed-EGFR back to the cell surface. 相似文献
20.
Fajka-Boja R Blaskó A Kovács-Sólyom F Szebeni GJ Tóth GK Monostori E 《Cellular and molecular life sciences : CMLS》2008,65(16):2586-2593
Mammalian galectin-1 (Gal-1), a beta-galactoside-binding lectin has a prominent role in regulating cell adhesion, cell growth and immune responses. Downregulation of these biological functions may occur via internalization of Gal-1. In the present study we have investigated the mechanism and possible mediator(s) of Gal-1 endocytosis. We show that internalization occurs at a temperature higher than 22 degrees C in an energy dependent fashion. After one hour incubation Gal-1 localizes in the Golgi system within the cells, and then disappears without accumulation in degradation compartments, such as lysosomes. Based on their strong intracellular co-localization, two glycoconjugates, GM1 ganglioside and CD7 are implicated in the sorting of internalized Gal-1 into Golgi. Other known Gal-1 binding glycoproteins on T cells (CD2, CD3, CD43 and CD45) do not cointernalize with the lectin. Internalization of Gal-1 depends on its lectin activity and follows dual pathways involving clathrin-coated vesicles and raft-dependent endocytosis. 相似文献