Journal of Cell Science
1477-9137
0021-9533
Anh Quốc
Cơ quản chủ quản: COMPANY BIOLOGISTS LTD , Company of Biologists Ltd
Lĩnh vực:
Cell Biology
Phân tích ảnh hưởng
Thông tin về tạp chí
Journal of Cell Science is committed to publishing the full range of topics in cell biology, and the single most important criterion for acceptance is scientific excellence. Articles must therefore pose and test a significant hypothesis that will provide novel perspectives and approaches to understanding cell biology, and will stimulate the interest of the broad readership of the journal.
Các bài báo tiêu biểu
Expression and localization of α-adaptin isoforms ABSTRACT
There are two α-adaptin genes, αA and αC, which in brain encode proteins of of Mr 108×103 and 104×103, respectively. Although both mRNAs can be detected on northern blots of brain and liver, the higher molecular mass polypeptide can only be detected on western blots of brain. Here we explain these observations by showing that αA is alternatively spliced and that the protein product in most tissues is different from the one expressed in brain in that it is missing 21 amino acids within the hinge region, giving it a similar mobility to that of αC. Monospecific antibodies were raised against the various α-adaptin isoforms and used to compare their distribution in cells and tissues. Both αA and αC are co-assembled into the same coated pits, and the larger isoform of αA is co-assembled with the smaller isoforms of α-adaptin, both in cells that naturally express it and in transfected cells. Examination of brain and spinal cord sections, labelled either for the larger isoform of αA or for αC, reveals that that the two are to some extent differentially distributed, consistent with previous in situ hybridisation studies. This finding, combined with the observation that there is considerable variability in the relative expression of the two isoforms in different tissues, indicates that the two genes are switched on in response to different stimuli. Moreover, the larger isoform of αA appears to be more efficiently concentrated in the nerve terminals than αC, which is found not only at the terminals but also diffusely distributed in the cell bodies and dendrites. This suggests that αC may play more of a role in the recycling of membrane components throughout the cell.
Tập 108 Số 8 - Trang 2865-2875 - 1995
Membrane remodeling in clathrin-mediated endocytosis ABSTRACT
Clathrin-mediated endocytosis is an essential cellular mechanism by which all eukaryotic cells regulate their plasma membrane composition to control processes ranging from cell signaling to adhesion, migration and morphogenesis. The formation of endocytic vesicles and tubules involves extensive protein-mediated remodeling of the plasma membrane that is organized in space and time by protein–protein and protein–phospholipid interactions. Recent studies combining high-resolution imaging with genetic manipulations of the endocytic machinery and with theoretical approaches have led to novel multifaceted phenomenological data of the temporal and spatial organization of the endocytic reaction. This gave rise to various – often conflicting – models as to how endocytic proteins and their association with lipids regulate the endocytic protein choreography to reshape the plasma membrane. In this Review, we discuss these findings in light of the hypothesis that endocytic membrane remodeling may be determined by an interplay between protein–protein interactions, the ability of proteins to generate and sense membrane curvature, and the ability of lipids to stabilize and reinforce the generated membrane shape through adopting their lateral distribution to the local membrane curvature.
Tập 131 Số 17 - 2018
The adaptor protein Dab2 sorts LDL receptors into coated pits independently of AP-2 and ARH Clathrin-mediated endocytosis requires cargo-specific adaptor proteins that recognize specific receptors and recruit them into coated pits. ARH [also called low-density lipoprotein receptor (LDLR) adaptor protein] serves as an adaptor for LDLR endocytosis in liver. However, ARH is dispensable for LDL uptake by some other cell types. Here, we show that the adaptor Dab2 plays a major role in LDLR internalization in HeLa cells and fibroblasts. Dab2 mediates internalization of LDLRs but not transferrin receptors independently of ARH and the classic clathrin adaptor AP-2. If Dab2 is absent, ARH can mediate LDLR endocytosis, but its action requires AP-2. Furthermore, the rate of LDLR endocytosis is decreased when Dab2 is absent and Dab2, but not ARH, catalyzes the efficient clustering of LDLR into coated pits. Dab2 activity requires its binding to clathrin, LDLR and phospholipids. Dab2 is also involved in moving LDLRs off filopodia. We suggest that Dab2 is a cargo-specific endocytic adaptor protein, stably associating with phospholipids and clathrin to sort LDLR to nascent-coated pits, whereas ARH might accelerate later steps in LDLR endocytosis in cooperation with AP-2.
Tập 119 Số 20 - Trang 4235-4246 - 2006
The mouse <i>brown</i> (<i>b</i>) locus protein has dopachrome tautomerase activity and is located in lysosomes in transfected fibroblasts ABSTRACT
Many genes mapping to pigmentation loci are involved in the regulation of melanin synthesis in the mouse. The brown (b) locus controls black/brown coat coloration, and its product has significant homology to the key melanogenic enzyme tyrosinase. This has led to suggestions that the b-protein is itself a melanogenic enzyme. In order to investigate its function, we have established lines of mouse fibroblasts stably expressing the b-protein by co-transfection of a b-protein expression vector and a plasmid conferring resistance to the antibiotic G418. The b-protein synthesised by these cells has the expected molecular mass of 75 kDa and reacts with three different anti-b-protein antibodies. We were unable to confirm previous reports that the b-protein has tyrosinase or catalase activity, but detected stereospecific dopachrome tautomerase activity in b-proteinexpressing fibroblasts. This dopachrome tautomerase binds to Concanavalin A-Sepharose, and the major product of its action on L-dopachrome is 5,6-di-hydroxyindole-2-carboxylic acid. Since this activity is not present in untransfected cells we conclude that the b-protein has dopachrome tautomerase activity. Fibroblasts do not contain melanosomes, the specialised organelles in which the b-protein is located in melanocytes. Nevertheless, indirect immunofluorescence localisation of the b-protein in transfected fibroblasts produces a distinctive pattern of intense juxtanuclear staining combined with punctate cytoplasmic staining. Double-labelling shows co-localisation of the b-protein with the late endosomal/lysosomal markers β-glucuronidase and LAMP-1, both in transfected fibroblasts and in mouse melanoma cells. These findings are consistent with the hypothesis that melanosomes are closely related to lysosomes.
Tập 106 Số 1 - Trang 153-166 - 1993
The mRNA-like noncoding RNA Gomafu constitutes a novel nuclear domain in a subset of neurons Recent transcriptome analyses have revealed that a large body of noncoding regions of mammalian genomes are actually transcribed into RNAs. Our understanding of the molecular features of these noncoding RNAs is far from complete. We have identified a novel mRNA-like noncoding gene, named Gomafu, which is expressed in a distinct set of neurons in the mouse nervous system. Interestingly, spliced mature Gomafu RNA is localized to the nucleus despite its mRNA-like characteristics, which usually act as potent export signals to the cytoplasm. Within the nucleus, Gomafu RNA is detected as numerous spots that do not colocalize with known nuclear domain markers. Gomafu RNA is extremely insoluble and remains intact after nuclear matrix preparation. Furthermore, heterokaryon assays revealed that Gomafu RNA does not shuttle between the nucleus and cytoplasm, but is retained in the nucleus after its transcription. We propose that Gomafu RNA represents a novel family of mRNA-like noncoding RNA that constitutes a cell-type-specific component of the nuclear matrix.
Tập 120 Số 15 - Trang 2498-2506 - 2007
Role of PPAR γ and EGFR signalling in the urothelial terminal differentiation programme Recently, considerable interest has focused on the ability of activated peroxisome proliferator-activated receptor γ (PPARγ) to promote cytodifferentiation in adipocytes and some carcinoma cells; however, the role of PPARγ in normal epithelial cytodifferentiation is unknown. Using uroplakin (UP) gene expression as a specific correlate of terminal urothelial cytodifferentiation, we investigated the differentiation-inducing effects of PPARγ activation in normal human urothelial (NHU) cells grown as finite cell lines in monoculture. Two high-affinity activators of PPARγ, troglitazone (TZ) and rosiglitazone (RZ) induced the expression of mRNA for UPII and UPIb and, to a lesser extent, UPIa. The specificity of the effect was shown by pretreating cells with a PPARγ antagonist, GW9662, which attenuated the TZ-induced response in a dose-specific manner. The PPARγ-mediated effect on UP gene expression was maximal when there was concurrent inhibition of autocrine-activated epidermal growth factor receptor (EGFR) signalling through either the phosphatidylinositol 3-kinase or extracellular signal-regulated kinase (ERK) pathways. The use of a specific EGFR tyrosine kinase inhibitor, PD153035, correlated with PPARγ dephosphorylation and translocation to the nucleus, indicating a mechanism for regulating the balance between proliferation and differentiation. This is the first identification of specific factors involved in regulating differentiation-associated gene changes in urothelium and the first unambiguous evidence of a role for PPARγ signalling in the terminal differentiation programme of a normal epithelium.
Tập 117 Số 10 - Trang 2029-2036 - 2004
Molecular cloning of a 47 kda tissue-specific and differentiationdependent urothelial cell surface glycoprotein ABSTRACT
Despite the fact that bladder epithelium has many interesting biological features and is a frequent site of carcinoma formation, relatively little is known about its biochemical differentiation. We have shown recently that a 47 kDa glycoprotein, uroplakin III (UPIII), in conjunction with uroplakins I (27 kDa) and II (15 kDa), forms the asymmetric unit membrane (AUM) - a highly specialized biomembrane characteristic of the apical surface of bladder epithelium. Deglycosylation and cDNA sequencing revealed that UPIII contains up to 20 kDa of N-linked sugars attached to a core protein of 28.9 kDa. The presence of an N-terminal signal peptide sequence and a single transmembrane domain located near the C terminus, plus the N-terminal location of all the potential N-glycosylation sites, points to a type I (N-exo/C-cyto) configuration. Thus the mass of the extracellular domain (20 kDa plus up to 20 kDa of sugar) of UPIII greatly exceeds that of its intracellular domain (5 kDa). Such an asymmetrical mass distribution, a feature shared by the other two major uroplakins, provides a molecular explanation as to why the luminal leaflet of AUM is almost twice as thick as the cytoplasmic one. The fact that of the three major proteins of AUM only UPIII has a significant cytoplasmic domain suggests that this molecule may play an important role in AUM-cytoskeleton interaction in terminally differentiated urothelial cells.
Tập 106 Số 1 - Trang 31-43 - 1993
The morphology of the granule cells of the olfactory bulb ABSTRACT
The granule cells of the olfactory bulb of the rat have been studied in material prepared by the Golgi-Kopsch method for examination with the light microscope, and in material examined with the electron microscope. With the Golgi method, the granule cells are found to have no process which can be identified as a typical axon, but from the superficial aspect of the somata stout peripheral processes arise and pass into the overlying external plexiform layer, while from the opposite side of the cell body several thinner deep dendrites extend towards the deeper parts of the bulb. Both types of processes, as well as the penkarya, have numerous spine-like appendages. On the distal portions of the peripheral processes in the external plexiform layer the appendages are much larger than on the deeper parts of the cell. The deep dendrites have localized swellings along their length which give them a varicose appearance, appendages often arise from these vancosities. The electron-microscopic features of the granule cells correspond well with the appearance of these cells in material impregnated with the Golgi method. The cell somata are small, with very little cytoplasm, and have a relatively large nucleus. The peripheral processes can be identified passing superficially from the penkarya of the granule cells; at their junction with the cell body their appearance is typically dendritic; all the cytoplasmic organelles found in the cytoplasm extend into these processes and none of the features of the initial segments of axons are found. In the external plexiform layer large spine-like appendages, which have been termed ‘gemmules’, arise from the distal portions of the peripheral processes, and participate in reciprocal synapses with the dendrites of mitral and tufted cells. The deep dendrites are much finer than the peripheral processes, and the vancosities which are seen in Golgi material may also be found with the electron microscope. Spines are found on all parts of the granule cells in the granule cell layer, including the peripheral processes, the penkarya and the deep dendrites. In addition to a spine apparatus, these spines commonly have numerous inclusions, including mitochondria, ribosomes, and vesicles which are the same size and shape as the synaptic vesicles present in the gemmules; no synapses oriented away from the spines have ever been found.
Tập 7 Số 1 - Trang 91-122 - 1970
The syndapin protein family: linking membrane trafficking with the cytoskeleton Syndapins – also called PACSINs – are highly conserved Src-homology 3 (SH3)-domain-containing proteins that seem to exist in all multicellular eukaryotes. They interact with the large GTPase dynamin and several other proteins implicated in vesicle trafficking. Syndapin-dynamin complexes appear to play an important role in vesicle fission at different donor membranes, including the plasma membrane (endocytosis) and Golgi membranes. In addition, syndapins are implicated in later steps of vesicle cycling in neuronal and non-neuronal cells. Syndapins also interact with N-WASP, a potent activator of the Arp2/3 complex that forms a critical part of the actin polymerization machinery. Syndapin oligomers can thereby couple bursts of actin polymerization with the vesicle fission step involving dynamins. This allows newly formed vesicles to move away from the donor membrane driven by actin polymerization. Syndapins also engage in additional interactions with molecules involved in several signal transduction pathways, producing crosstalk at the interface between membrane trafficking and the cytoskeleton. Given the distinct expression patterns of the different syndapins and their splice forms, these proteins could have isoform-specific functions.
Tập 117 Số 15 - Trang 3077-3086 - 2004
Potential role of MCP-1 in endothelial cell tight junction `opening': signaling via Rho and Rho kinase The expression of the monocyte chemoattractant protein-1 (MCP-1) receptor CCR2 by brain endothelial cells suggests that MCP-1 may have other functions than purely driving leukocyte migration into brain parenchyma during inflammation. This study examines one of these potential novel roles of MCP-1 regulation of endothelial permeability using primary cultures of mouse brain endothelial cells. MCP-1 induces reorganization of actin cytoskeleton (stress fiber formation) and redistribution of tight junction proteins, ZO-1, ZO-2 occludin and claudin-5, from the Triton X-100-soluble to the Triton X-100-insoluble fractions. These morphological changes are associated with a decrease in transendothelial electrical membrane resistance and an increase in [14C]inulin permeability. MCP-1 did not induce these events in brain endothelial cells prepared from mice genotype CCR2–/–. The Rho kinase inhibitor Y27632 and inhibition of Rho (C3 exoenzyme, and dominant negative mutant of Rho, RhoT19N) prevented MCP-1-induced stress fiber assembly, reorganization of tight junction proteins and alterations in endothelial permeability. In all, this suggests that a small GTPase Rho and Rho kinase have a pivotal role in MCP-1-induced junction disarrangement. These data are the first to strongly suggest that MCP-1, via CCR2 present on brain endothelial cells, contributes to increased brain endothelial permeability.
Tập 116 Số 22 - Trang 4615-4628 - 2003