p115 is a general vesicular transport factor related to the yeast endoplasmic reticulum to Golgi transport factor Uso1p.

Sapperstein, Stephanie K.; Walter, David M.; Grosvenor, A R; Heuser, John E.; Waters, M. Gerard

doi:10.1073/pnas.92.2.522

Cited by 185 publications

(160 citation statements)

References 59 publications

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“…In this study, we used the N-terminal cytoplasmic domain of IRAP, residues 1-109, conjugated to a chitin-binding protein in order to find cytosolic proteins involved in GSV trafficking, and we identified p115 as one such protein. p115 is known to be involved in vesicular traffic, for example, by tethering vesicle in the process of docking and fusing in the Golgi apparatus and in in vitro membrane trafficking protocols (Waters et al, 1992;Barroso et al, 1995;Sapperstein et al, 1995). Here, we document that p115 binds GLUT4 storage vesicles via IRAP and plays an important role in GLUT4 trafficking.…”

mentioning

confidence: 68%

“…In vitro vesicle fusion assays suggest p115 to be a general factor required for transport vesicle fusion to target membranes (Barroso et al, 1995;Sapperstein et al, 1995), in the present example, presumably the plasma membrane. And although the intensity of p115 fluorescence shows it to be mainly perinuclear and similar to syntaxin 6 and GM130 in adipocytes ( Figure 5), subcellular fractionation reveals it to be ubiquitous in adipocytes membrane compartments, including the PM, as well as in the cytosol ( Figure 2).…”

Section: Discussionmentioning

confidence: 98%

“…We then passed cytosol from rat adipocytes and 3T3-L1 cultured adipocytes over an IRAP-CBD affinity column and found a number of Coomassie staining bands that could be specifically eluted with IRAP 1-109 peptide (unpublished data). The slowest migrating band from both rat and mouse cytosols was subjected to proteomic analysis by mass spectrometry (Dr. W. Lane, Harvard Microchemistry and Proteomics Facility, Boston, MA), and nine individual peptides were identified corresponding to p115 (Waters et al, 1992;Barroso et al, 1995;Sapperstein et al, 1995). We confirmed this identification by Western blotting as shown in Figure 1 (see also Figure 4) where we passed cytosol from 3T3-L1 adipocytes over the affinity column and see nearly complete retention of p115 on an IRAP-CBD column and specific elution with IRAP 1-109 peptide.…”

Section: Identification Of P115 As An Irap-binding Proteinmentioning

confidence: 99%

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p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

Hosaka

Brooks

Presman

et al. 2005

MBoC

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Insulin-regulated aminopeptidase (IRAP) is an abundant cargo protein of Glut4 storage vesicles (GSVs) that traffics to and from the plasma membrane in response to insulin. We used the amino terminus cytoplasmic domain of IRAP, residues 1-109, as an affinity reagent to identify cytosolic proteins that might be involved in GSV trafficking. In this way, we identified p115, a peripheral membrane protein known to be involved in membrane trafficking. In murine adipocytes, we determined that p115 was localized to the perinuclear region by immunofluorescence and throughout the cell by fractionation. By immunofluorescence, p115 partially colocalizes with GLUT4 and IRAP in the perinuclear region of cultured fat cells. The amino terminus of p115 binds to IRAP and overexpression of a N-terminal construct results in its colocalization with GLUT4 throughout the cell. Insulin-stimulated GLUT4 translocation is completely inhibited under these conditions. Overexpression of p115 C-terminus has no significant effect on GLUT4 distribution and translocation. Finally, expression of the p115 N-terminus construct has no effect on the distribution and trafficking of GLUT1. These data suggest that p115 has an important and specific role in insulin-stimulated Glut4 translocation, probably by way of tethering insulin-sensitive Glut4 vesicles at an as yet unknown intracellular site. INTRODUCTIONInsulin normalizes blood glucose levels by mobilizing the muscle and adipocyte glucose transporter isoform, GLUT4, from intracellular storage vesicles and moving it to the plasma membrane (Simpson et al., 2001;Bryant et al., 2002). Various models of GLUT4 trafficking suggest that GLUT4 must exist in more than one intracellular compartment and the major insulin sensitive pool is localized to a compartment that is distinct from endosomal markers and is commonly referred to as glucose transporter storage vesicles (GSVs), or insulin responsive vesicle (IRVs). Despite the critical function of glucose transport in glucose homeostasis, many of the details by which adipocytes and muscle form a pathway of insulin-sensitive GLUT4 trafficking remain unknown. However, it is virtually certain that the major cargo proteins of GSVs must interact with a number of cytosolic and membrane proteins, such as adaptors and tethers, in order to be properly sorted and regulated by insulin.Insulin-responsive aminopeptidase (IRAP) was identified as an abundant cargo protein associated with GLUT4 vesicles that translocates in response to insulin in a manner seemingly identical to GLUT4 Mastick et al., 1994;Keller et al., 1995;Malide et al., 1997;Martin et al., 1997;Ross et al., 1997). In fact, it is more abundantly expressed in vesicles than the transporter (Kupriyanova et al., 2002). When the cytoplasmic N-terminus of IRAP was microinjected into 3T3-L1 adipocytes, GLUT4 was localized on the plasma membrane even in the basal state (Waters et al., 1997), suggesting IRAP can play a role in GSV movement/targeting. A chimeric protein containing the intracellular domain of IRAP and ...

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mentioning

confidence: 68%

Section: Discussionmentioning

confidence: 98%

Section: Identification Of P115 As An Irap-binding Proteinmentioning

confidence: 99%

See 1 more Smart Citation

p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

Hosaka

Brooks

Presman

et al. 2005

MBoC

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“…Homodimeric p115 contains two N-terminal globular head domains linked by a central coiled-coil domain (20). The extreme C termini form an acidic tail domain that mediates p115 binding to giantin and GM130 (21,22).…”

Section: Resultsmentioning

confidence: 99%

Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis

Puthenveedu

Linstedt

2004

Proc. Natl. Acad. Sci. U.S.A.

124

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Functional characterization of protein interactions in mammalian systems has been hindered by the inability to perform complementation analyses in vivo. Here, we use functional replacement of the vesicle docking protein p115 to separate its essential from its nonessential interactions. p115 is required for biogenesis of the Golgi apparatus, but it is unclear whether its mechanism of action requires its golgin and͞or SNARE interactions. Short interfering RNA-mediated knockdown of p115 induced extensive Golgi fragmentation and impaired secretory traffic. Reassembly of a structurally and functionally normal Golgi occurred on expression of a p115 homologue not recognized by the short interfering RNA. Strikingly, versions of p115 lacking its phosphorylation site and the golgin-binding domains also restored the Golgi apparatus in cells lacking endogenous p115. In contrast, the p115 SNARE-interacting domain was required for Golgi biogenesis. This suggests that p115 acts directly, rather than via a tether, to catalyze trans-SNARE complex formation preceding membrane fusion.Golgi apparatus ͉ RNA interference ͉ giantin ͉ GM130

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“…The prototype of this group of proteins is p115, but other golgins assume analogous conformations as well. 21,22 It is interesting to note that in some of these proteins one extension is associated with microtubules via dyneins. 23,24 Microtubules and dynein, however, do not appear to be required for prApe1 transport.…”

Section: Addendummentioning

confidence: 99%

Atg11 Directs Autophagosome Cargoes to the PAS Along Actin Cables

Monastyrska

Shintani²,

Klionsky³

et al. 2005

Autophagy

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p115 is a general vesicular transport factor related to the yeast endoplasmic reticulum to Golgi transport factor Uso1p.

Cited by 185 publications

References 59 publications

p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis

Atg11 Directs Autophagosome Cargoes to the PAS Along Actin Cables

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