The cytoplasmic domain mediates localization of furin to the trans-Golgi network en route to the endosomal/lysosomal system.

Bosshart, Herbert; Humphrey, Jeffrey S.; Deignan, E; Davidson, Jesse; Drazba, Judith A.; Yuan, Lydia C.; Oorschot, Viola; Peters, Peter J.; Bonifacino, Juan S.

doi:10.1083/jcb.126.5.1157

Cited by 162 publications

(206 citation statements)

References 62 publications

(90 reference statements)

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“…A SmaI/XhoI restriction fragment containing the fulllength receptor was ligated into pCIneo expression vector. pCDM8.1 expression vector (Invitrogen) encoding the interleukin-2 receptor ␣-subunit (tac) fused in frame with the transmembrane and cytoplasmic domain of furin (tac-furin) was kindly provided by Dr. Juan Bonifacino (19). The pEYFP-Golgi vector encoding the enhanced yellow fluorescent protein fused to the N-terminal 81 amino acids of the precursor to human ␤-1,4 galactosyltransferase was obtained from CLONTECH.…”

Section: Methodsmentioning

confidence: 99%

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Identification and Characterization of SNX15, a Novel Sorting Nexin Involved in Protein Trafficking

Phillips¹,

Barr²,

Haft³

et al. 2001

Journal of Biological Chemistry

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Sorting nexins are a family of phox homology domain containing proteins that are homologous to yeast proteins involved in protein trafficking. We have identified a novel 342-amino acid residue sorting nexin, SNX15, and a 252-amino acid splice variant, SNX15A. Unlike many sorting nexins, a SNX15 ortholog has not been identified in yeast or Caenorhabditis elegans. By Northern blot analysis, SNX15 mRNA is widely expressed. Although predicted to be a soluble protein, both endogenous and overexpressed SNX15 are found on membranes and in the cytosol. The phox homology domain of SNX15 is required for its membrane association and for association with the platelet-derived growth factor receptor. We did not detect association of SNX15 with receptors for epidermal growth factor or insulin. However, overexpression of SNX15 led to a decrease in the processing of insulin and hepatocyte growth factor receptors to their mature subunits. Immunofluorescence studies showed that SNX15 overexpression resulted in mislocalization of furin, the endoprotease responsible for cleavage of insulin and hepatocyte growth factor receptors. Based on our data and the existing findings with yeast orthologs of other sorting nexins, we propose that overexpression of SNX15 disrupts the normal trafficking of proteins from the plasma membrane to recycling endosomes or the trans-Golgi network.Intracellular vesicle traffic in both yeast and mammals requires the function of numerous proteins that mediate multiple processes including cargo selection, vesicle budding, and fusion of vesicles with specific targets (1-3). Sorting nexins (SNXs) 1 are a family of widely expressed proteins believed to be part of the complex molecular machinery required for protein trafficking (4). Mammalian SNXs are homologous to several yeast proteins (e.g. Vps5p, Mvp1p, and Grd19p) for which there is strong genetic evidence demonstrating a role in protein trafficking (5-7). SNX1 was identified using the yeast two-hybrid system by virtue of its ability to bind to the cytoplasmic domain of the epidermal growth factor (EGF) receptor (8). Subsequently, SNX1 was shown to also interact with receptors for insulin, platelet-derived growth factor (PDGF), transferrin, and leptin (4). Furthermore, SNX1 is the mammalian ortholog of Vps5p, a yeast protein that is a component of a multimeric complex (termed the "retromer complex") involved in retrograde transport of proteins from prevacuolar endosomes to the TGN (9). We have previously described three additional sorting nexins: SNX2, SNX3, and SNX4 (4); and 10 additional sorting nexins (SNX5-14) have been deposited in GenBank TM . Like SNX1, both SNX2 and SNX4 associate with various receptors. In addition, SNX1, SNX2, and SNX4 assemble into oligomeric structures (4). All 14 SNX molecules contain a phox homology (PX) domain, a conserved sequence of unknown function first identified in the p40 phox and p47 phox subunits of the NADPH oxidase complex (10). PX domains consist of ϳ100 amino acid residues, and most contain a proline-rich sequence ...

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Section: Methodsmentioning

confidence: 99%

“…The chimera consisted of the extracellular domain of the ␣ subunit of the interleukin-2 receptor fused to the transmembrane and cytoplasmic domains of furin (19). It has previously been shown that both native furin and the tac-furin chimera are localized predominantly in the TGN.…”

Section: Fig 11 Expression Of Snx15 Impairs Post-translational Procmentioning

confidence: 99%

Identification and Characterization of SNX15, a Novel Sorting Nexin Involved in Protein Trafficking

Phillips¹,

Barr²,

Haft³

et al. 2001

Journal of Biological Chemistry

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“…The molecular mass of the AP-4 complex was calculated on the basis of its Stokes radius and sedimentation coefficient, assuming a partial specific volume of 0.72-0.75 cm 3 /g (24). Immunofluorescence Microscopy-HeLa cell monolayers were grown on glass coverslips and in some cases treated with brefeldin A or transiently transfected with HA epitope-tagged constructs of TGN38 or furin (7,25). Cells were fixed in methanol/acetone (1:1, v/v) for 10 min at Ϫ20°C and subsequently air-dried.…”

Section: Methodsmentioning

confidence: 99%

AP-4, a Novel Protein Complex Related to Clathrin Adaptors

Dell’Angelica

Mullins²,

Bonifacino

1999

Journal of Biological Chemistry

270

199

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Here we report the identification and characterization of AP-4, a novel protein complex related to the heterotetrameric AP-1, AP-2, and AP-3 adaptors that mediate protein sorting in the endocytic and late secretory pathways. The key to the identification of this complex was the cloning and sequencing of two widely expressed, mammalian cDNAs encoding new homologs of the adaptor ␤ and subunits named ␤4 and 4, respectively. An antibody to ␤4 recognized in human cells an ϳ83-kDa polypeptide that exists in both soluble and membrane-associated forms. Gel filtration, sedimentation velocity, and immunoprecipitation experiments revealed that ␤4 is a component of a multisubunit complex (AP-4) that also contains the 4 polypeptide and two additional adaptor subunit homologs named 4 (-ARP2) and ⑀. Immunofluorescence analyses showed that AP-4 is associated with the trans-Golgi network or an adjacent structure and that this association is sensitive to the drug brefeldin A. We propose that, like the related AP-1, AP-2, and AP-3 complexes, AP-4 plays a role in signal-mediated trafficking of integral membrane proteins in mammalian cells.

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“…5). Localization of furin to the TGN requires a bipartite motif that is composed of the casein kinase 2 (CK2)-phosphorylated acidic cluster (EECPpSDpSEEDE, where p highlights the serine residues that are phosphorylated) and a membrane-proximal segment containing two hydrophobic motifs (YKGL and LI) [23][24][25][26][27] (FIG. 5).…”

Section: Localization To the Trans-golgi Networkmentioning

confidence: 99%

Furin at the cutting edge: From protein traffic to embryogenesis and disease

Thomas

2002

Nat Rev Mol Cell Biol

1,100

1,285

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Furin catalyses a simple biochemical reaction -the proteolytic maturation of proprotein substrates in the secretory pathway. But the simplicity of this reaction belies furin's broad and important roles in homeostasis, as well as in diseases ranging from Alzheimer's disease and cancer to anthrax and Ebola fever. This review summarizes various features of furin -its structural and enzymatic properties, intracellular localization, trafficking, substrates, and roles in vivo.Furin is a cellular ENDOPROTEASE that was identified in 1990 (BOX 1); it proteolytically activates large numbers of PROPROTEIN substrates in secretory pathway compartments. As well as activating pathogenic agents (BOX 2), furin has an essential role in embryogenesis, and catalyses the maturation of a strikingly diverse collection of proprotein substrates. These range from growth factors and receptors to extracellular-matrix proteins and even other protease systems that control disease. Until recently, furin was thought to be an unglamorous housekeeping protein; however, furin's crucial role in so many different cellular events -and in diseases ranging from from anthrax and bird flu (BOX 2) to cancer, dementia and Ebola fever -has caused researchers to re-evaluate it. In this review, I summarize the various features of furin: its structural and enzymatic properties, autoactivation, intracellular localization and trafficking; its substrates; and its roles in vivo, including the requirement for furin in determining the PATHOGENICITY of many viruses and bacteria.

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The cytoplasmic domain mediates localization of furin to the trans-Golgi network en route to the endosomal/lysosomal system.

Cited by 162 publications

References 62 publications

Identification and Characterization of SNX15, a Novel Sorting Nexin Involved in Protein Trafficking

Identification and Characterization of SNX15, a Novel Sorting Nexin Involved in Protein Trafficking

AP-4, a Novel Protein Complex Related to Clathrin Adaptors

Furin at the cutting edge: From protein traffic to embryogenesis and disease

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