Vesicle Formation at the Plasma Membrane and Trans-Golgi Network: The Same but Different

A crucial step in lysosomal biogenesis is catalyzed by “uncovering” enzyme (UCE), which removes a coveringN-acetylglucosamine from the mannose 6-phosphate (Man-6-P) recognition marker on lysosomal hydrolases. This study shows that UCE resides in the trans-Golgi network (TGN) and cycles between the TGN and plasma membrane. The cytosolic domain of UCE contains two potential endocytosis motifs: 488YHPL and C-terminal 511NPFKD. YHPL is shown to be the more potent of the two in retrieval of UCE from the plasma membrane. A green-fluorescent protein-UCE transmembrane-cytosolic domain fusion protein colocalizes with TGN 46, as does endogenous UCE in HeLa cells, showing that the transmembrane and cytosolic domains determine intracellular location. These data imply that the Man-6-P recognition marker is formed in the TGN, the compartment where Man-6-P receptors bind cargo and are packaged into clathrin-coated vesicles.

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Identification of a di-leucine motif within the C terminus domain of the Menkes disease protein that mediates endocytosis from the plasma membrane

Journal of Cell Science ◽

10.1242/jcs.112.11.1721 ◽

1999 ◽

Vol 112 (11) ◽

pp. 1721-1732 ◽

Cited By ~ 2

Author(s):

M.J. Francis ◽

E.E. Jones ◽

E.R. Levy ◽

R.L. Martin ◽

S. Ponnambalam ◽

...

Keyword(s):

Plasma Membrane ◽

Golgi Apparatus ◽

Transmembrane Domain ◽

Menkes Disease ◽

Cytoplasmic Domain ◽

Endocytic Pathway ◽

Site Directed Mutagenesis ◽

Trans Golgi Network ◽

C Terminus ◽

Golgi Network

The protein encoded by the Menkes disease gene (MNK) is localised to the Golgi apparatus and cycles between the trans-Golgi network and the plasma membrane in cultured cells on addition and removal of copper to the growth medium. This suggests that MNK protein contains active signals that are involved in the retention of the protein to the trans-Golgi network and retrieval of the protein from the plasma membrane. Previous studies have identified a signal involved in Golgi retention within transmembrane domain 3 of MNK. To identify a motif sufficient for retrieval of MNK from the plasma membrane, we analysed the cytoplasmic domain, downstream of transmembrane domain 7 and 8. Chimeric constructs containing this cytoplasmic domain fused to the reporter molecule CD8 localised the retrieval signal(s) to 62 amino acids at the C terminus. Further studies were performed on putative internalisation motifs, using site-directed mutagenesis, protein expression, chemical treatment and immunofluorescence. We observed that a di-leucine motif (L1487L1488) was essential for rapid internalisation of chimeric CD8 proteins and the full-length Menkes cDNA from the plasma membrane. We suggest that this motif mediates the retrieval of MNK from the plasma membrane into the endocytic pathway, via the recycling endosomes, but is not sufficient on its own to return the protein to the Golgi apparatus. These studies provide a basis with which to identify other motifs important in the sorting and delivery of MNK from the plasma membrane to the Golgi apparatus.

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Ride the wave: Retrograde trafficking becomes Ca2+ dependent with BAIAP3

The Journal of Cell Biology ◽

10.1083/jcb.201706007 ◽

2017 ◽

Vol 216 (7) ◽

pp. 1887-1889 ◽

Cited By ~ 3

Author(s):

Jakob B. Sørensen

Keyword(s):

Angiogenesis Inhibitor ◽

Secretory Vesicle ◽

Vesicle Formation ◽

Retrograde Trafficking ◽

Trans Golgi Network ◽

Cell Biol ◽

Golgi Network

The functions of four of the five proteins in the mammalian uncoordinated-13 (Munc13) family have been identified as priming factors in SNARE-dependent exocytosis. In this issue, Zhang et al. (2017. J. Cell Biol. https://doi.org/10.1083/jcb.201702099) show that the fifth member, BAIAP3 (brain-specific angiogenesis inhibitor I–associated protein 3), acts in retrograde trafficking by returning secretory vesicle material to the trans-Golgi network. In its absence, secretory vesicle formation is impaired, leading to accumulation of immature vesicles, or lysosomal vesicle degradation.

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The dynamin-cortactin complex as a mediator of vesicle formation at the trans-Golgi network

The Golgi Apparatus ◽

10.1007/978-3-211-76310-0_18 ◽

2009 ◽

pp. 301-313

Author(s):

Shaun Weller ◽

Hong Cao ◽

Mark A. McNiven

Keyword(s):

Vesicle Formation ◽

Trans Golgi Network ◽

Golgi Network

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Dual-color visualization of trans-Golgi network to plasma membrane traffic along microtubules in living cells

Journal of Cell Science ◽

10.1242/jcs.112.1.21 ◽

1999 ◽

Vol 112 (1) ◽

pp. 21-33 ◽

Cited By ~ 17

Author(s):

D. Toomre ◽

P. Keller ◽

J. White ◽

J.C. Olivo ◽

K. Simons

Keyword(s):

Plasma Membrane ◽

Protein Trafficking ◽

Membrane Traffic ◽

Living Cells ◽

Tubular Structures ◽

Trans Golgi Network ◽

Vesicular Stomatitis ◽

Golgi Network ◽

Color Visualization

The mechanisms and carriers responsible for exocytic protein trafficking between the trans-Golgi network (TGN) and the plasma membrane remain unclear. To investigate the dynamics of TGN-to-plasma membrane traffic and role of the cytoskeleton in these processes we transfected cells with a GFP-fusion protein, vesicular stomatitis virus G protein tagged with GFP (VSVG3-GFP). After using temperature shifts to block VSVG3-GFP in the endoplasmic reticulum and subsequently accumulate it in the TGN, dynamics of TGN-to-plasma membrane transport were visualized in real time by confocal and video microscopy. Both small vesicles (<250 nm) and larger vesicular-tubular structures (>1.5 microm long) are used as transport containers (TCs). These TCs rapidly moved out of the Golgi along curvilinear paths with average speeds of approximately 0.7 micrometer/second. Automatic computer tracking objectively determined the dynamics of different carriers. Fission and fusion of TCs were observed, suggesting that these late exocytic processes are highly interactive. To directly determine the role of microtubules in post-Golgi traffic, rhodamine-tubulin was microinjected and both labeled cargo and microtubules were simultaneously visualized in living cells. These studies demonstrated that exocytic cargo moves along microtubule tracks and reveals that carriers are capable of switching between tracks.

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AGS3-dependent trans-Golgi network membrane trafficking is essential for compaction in mouse embryos

Journal of Cell Science ◽

10.1242/jcs.243238 ◽

2020 ◽

Vol 133 (23) ◽

pp. jcs243238

Author(s):

Zheng-Wen Nie ◽

Ying-Jie Niu ◽

Wenjun Zhou ◽

Dong-Jie Zhou ◽

Ju-Yeon Kim ◽

...

Keyword(s):

Plasma Membrane ◽

Membrane Trafficking ◽

Fluorescent Protein ◽

Mouse Embryos ◽

Cell Contact ◽

Protein Signaling ◽

Cell Stage ◽

Trans Golgi Network ◽

Early Mouse ◽

Golgi Network

ABSTRACTActivator of G-protein signaling 3 (AGS3, also known as GPSM1) regulates the trans-Golgi network. The AGS3 GoLoco motif binds to Gαi and thereby regulates the transport of proteins to the plasma membrane. Compaction of early embryos is based on the accumulation of E-cadherin (Cdh1) at cell-contacted membranes. However, how AGS3 regulates the transport of Cdh1 to the plasma membrane remains undetermined. To investigate this, AGS3 was knocked out using the Cas9-sgRNA system. Both trans-Golgi network protein 46 (TGN46, also known as TGOLN2) and transmembrane p24-trafficking protein 7 (TMED7) were tracked in early mouse embryos by tagging these proteins with a fluorescent protein label. We observed that the majority of the AGS3-edited embryos were developmentally arrested and were fragmented after the four-cell stage, exhibiting decreased accumulation of Cdh1 at the membrane. The trans-Golgi network and TMED7-positive vesicles were also dispersed and were not polarized near the membrane. Additionally, increased Gαi1 (encoded by GNAI1) expression could rescue AGS3-overexpressed embryos. In conclusion, AGS3 reinforces the dynamics of the trans-Golgi network and the transport of TMED7-positive cargo containing Cdh1 to the cell-contact surface during early mouse embryo development.

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Yeast synaptobrevin, Snc1, engages distinct routes of postendocytic recycling mediated by a sorting nexin, Rcy1-COPI, and retromer

Molecular Biology of the Cell ◽

10.1091/mbc.e19-05-0290 ◽

2020 ◽

Vol 31 (9) ◽

pp. 944-962

Author(s):

Jordan T. Best ◽

Peng Xu ◽

Jack G. McGuire ◽

Shannon N. Leahy ◽

Todd R. Graham

Keyword(s):

Plasma Membrane ◽

Multiple Modes ◽

Sorting Nexin ◽

Trans Golgi Network ◽

Golgi Network

The yeast synaptobrevin, Snc1, uses multiple modes of postendocytic recycling to facilitate its return to either the trans-Golgi network or the plasma membrane. Snc1 primarily recycles via pathways dependent on either Rcy1/COPI or Snx4, while a smaller portion of the SNARE appears to be capable of being retrieved by retromer.

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Calneurons provide a calcium threshold for trans-Golgi network to plasma membrane trafficking

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0903001106 ◽

2009 ◽

Vol 106 (22) ◽

pp. 9093-9098 ◽

Cited By ~ 43

Author(s):

M. Mikhaylova ◽

P. P. Reddy ◽

T. Munsch ◽

P. Landgraf ◽

S. K. Suman ◽

...

Keyword(s):

Plasma Membrane ◽

Membrane Trafficking ◽

Trans Golgi Network ◽

Golgi Network

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The complex interactions of Chs5p, the ChAPs, and the cargo Chs3p

Molecular Biology of the Cell ◽

10.1091/mbc.e11-12-1015 ◽

2012 ◽

Vol 23 (22) ◽

pp. 4402-4415 ◽

Cited By ~ 15

Author(s):

Uli Rockenbauch ◽

Alicja M. Ritz ◽

Carlos Sacristan ◽

Cesar Roncero ◽

Anne Spang

Keyword(s):

Cell Cycle ◽

Plasma Membrane ◽

Chitin Synthase ◽

Tetratricopeptide Repeats ◽

Trans Golgi Network ◽

Complex Interactions ◽

C Terminus ◽

Bud Neck ◽

Domain Switch ◽

Golgi Network

The exomer complex is a putative vesicle coat required for the direct transport of a subset of cargoes from the trans-Golgi network (TGN) to the plasma membrane. Exomer comprises Chs5p and the ChAPs family of proteins (Chs6p, Bud7p, Bch1p, and Bch2p), which are believed to act as cargo receptors. In particular, Chs6p is required for the transport of the chitin synthase Chs3p to the bud neck. However, how the ChAPs associate with Chs5p and recognize cargo is not well understood. Using domain-switch chimeras of Chs6p and Bch2p, we show that four tetratricopeptide repeats (TPRs) are involved in interaction with Chs5p. Because these roles are conserved among the ChAPs, the TPRs are interchangeable among different ChAP proteins. In contrast, the N-terminal and the central parts of the ChAPs contribute to cargo specificity. Although the entire N-terminal domain of Chs6p is required for Chs3p export at all cell cycle stages, the central part seems to predominantly favor Chs3p export in small-budded cells. The cargo Chs3p probably also uses a complex motif for the interaction with Chs6, as the C-terminus of Chs3p interacts with Chs6p and is necessary, but not sufficient, for TGN export.

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