Immunolocalization of the cellular src protein in interphase and mitotic NIH c-src overexpresser cells.

David‐Pfeuty, Thérèse; Nouvian-Dooghe, Yolande

doi:10.1083/jcb.111.6.3097

Cited by 129 publications

(99 citation statements)

References 64 publications

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“…In line with this these results, we have found that a subpopulation of Fyn-GFP localizes with internalized EGF on endosomes, and this colocalization is more evident after 20 min of internalization. Consistently, Src, another member of the SFKs, also localizes partially to endosomes (55,61,62). Therefore, the three proteins that have been described to interact with the C-terminal region of Tom1L1, namely Fyn, Grb2, and p85, are known to be translocated to endosomes after EGF receptor internalization.…”

Section: Discussionmentioning

confidence: 79%

Interactions of TOM1L1 with the Multivesicular Body Sorting Machinery

Puertollano

2005

Journal of Biological Chemistry

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Tom1L1 (Tom1-like1) and related proteins Tom1 (Target of Myb1) and Tom1L2 (Tom1-like2) constitute a new protein family characterized by the presence of a VHS (Vps27p/Hrs/Stam) domain in the N-terminal portion followed by a GAT (GGA and Tom) domain. Recently it was demonstrated that the GAT domain of both Tom1 and Tom1L1 binds ubiquitin, suggesting that these proteins might participate in the sorting of ubiquitinated proteins into multivesicular bodies (MVBs). Here we report a novel interaction between Tom1L1 and members of the MVB sorting machinery. Specifically, we found that the VHS domain of Tom1L1 interacts with Hrs (Hepatocyte growth factor-regulated tyrosine kinase substrate), whereas a PTAP motif, located between the VHS and GAT domain of Tom1L1, is responsible for binding to TSG101 (tumor susceptibility gene 101). Myc epitopetagged Tom1L1 showed a cytosolic distribution but was recruited to endosomes following Hrs expression. In addition, Tom1L1 possesses several tyrosine motifs at the C-terminal region that mediate interactions with members of the Src family kinases and other signaling proteins such as Grb2 and p85. We showed that a fraction of Fyn kinase localizes at endosomes and that this distribution becomes more evident after epidermal growth factor internalization. Moreover, expression of a constitutive active form of Fyn also promoted the recruitment of Tom1L1 to enlarged endosomes. Taken together, we propose that Tom1L1 could act as an intermediary between signaling and degradative pathways.Regulated degradation of cell surface proteins, particularly receptors involved in signaling pathways, is essential for the control of many biological processes, including cell proliferation, survival, migration, and differentiation (1, 2). Internalized membrane-protein cargo can be delivered either to recycling vesicles for transport back to the cell surface or to late endosomes for transfer to lysosomes and degradation. Multivesicular bodies (MVBs) 1 are the compartments where this sorting event takes place (3). Thus, proteins to be degraded are included into small vesicles that invaginate into the lumen of the endosome. Subsequent fusion of MVBs with lysosomes releases these vesicles into the lysosome lumen, where they are degraded by lysosomal hydrolases. In contrast, proteins that remain in the limiting membrane of MVBs are recycled to the trans-Golgi network (TGN) or plasma membrane. In the last few years, a number of genetic and biochemical studies have allowed the characterization of the molecular machinery implicated in the formation of MVBs. Hrs appears to be the first protein to be recruited to endosomes (4 -6) through an interaction of its FIVE domain with phosphatidylinositol 3-phosphate (7), a lipid highly enriched in endosomal membranes (8). This is followed by the consecutive recruitment of three multisubunit complexes termed ESCRT-I (9), ESCRT-II (10), and ESCRT-III (11). Finally, the AAA-type ATPase Vps4 binds ESCRT-III and, following MVB vesicle formation, promotes the dissociation of the E...

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Section: Discussionmentioning

confidence: 79%

Interactions of TOM1L1 with the Multivesicular Body Sorting Machinery

Puertollano

2005

Journal of Biological Chemistry

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“…S1 D and E). Thus, it is attractive to predict that activation of a Golgi-associated Src kinase (22,23), when activated during the transit of nascent secretory proteins from the ER to the Golgi (3), could induce TGN vesiculation.…”

Section: Modulation Of Src Activity Affects Golgi Compartment Integrimentioning

confidence: 99%

Src kinase regulates the integrity and function of the Golgi apparatus via activation of dynamin 2

Weller

Capitani²,

Cao

et al. 2010

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

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The size and integrity of the Golgi apparatus is maintained via a tightly controlled regulation of membrane traffic using a variety of different signaling and cytoskeletal proteins. We have recently observed that activation of c-Src has profound effects on Golgi structure, leading to dramatically vesiculated cisternae in a variety of cell types. As the large GTPase dynamin (Dyn2) has been implicated in Golgi vesiculation during secretion, we tested whether inhibiting Dyn2 activity by expression of a Dyn2K44A mutant or siRNA knockdown could attenuate active Src-induced Golgi fragmentation. Indeed, these perturbations attenuated fragmentation, and expression of a Dyn2Y(231/597)F mutant protein that cannot be phosphorylated by Src kinase had a similar effect . Finally, we find that Dyn2 is markedly phosphorylated during the transit of VSV-G protein through the TGN whereas expression of the Dyn2Y(231/597)F mutant significantly reduces exit of the nascent protein from this compartment. These findings demonstrate that activation of Dyn2 by Src kinase regulates Golgi integrity and vesiculation during the secretory process.

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“…1 and 2). A significant fraction of Src is localized to the perinuclear region and the secretory vesicles in neuronal cells (3,4). It interacts with and phosphorylates proteins such as dynamin and synapsin (5-7) and synaptophysin (8) or cellugyrin (9), suggesting a role in the regulation of membrane traffic.…”

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confidence: 99%

Src Regulates Golgi Structure and KDEL Receptor-dependent Retrograde Transport to the Endoplasmic Reticulum

Bard

Mazelin

Péchoux-Longin

et al. 2003

Journal of Biological Chemistry

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The tyrosine kinase Src is present on the Golgi membranes. Its role, however, in the overall function and organization of the Golgi apparatus is unclear. We have found that in a cell line called SYF, which lacks the three ubiquitous Src-like kinases (Src, Yes, and Fyn), the organization of the Golgi apparatus is perturbed. The Golgi apparatus is composed of collapsed stacks and bloated cisternae in these cells. Expression of an activated form of Src relocated the KDEL receptor (KDEL-R) from the Golgi apparatus to the endoplasmic reticulum. Other Golgi-specific marker proteins were not affected under these conditions. Because of the specific effect of Src on the location of KDEL-R, we tested whether protein transport between ER and the Golgi apparatus involves Src. Transport of Pseudomonas exotoxin, which is transported to the ER by binding to the KDEL-R is accelerated by inhibition or genetic ablation of Src. Protein transport from ER to the Golgi apparatus however, is unaffected by Src deletion or inhibition. We propose that Src has an appreciable role in the organization of the Golgi apparatus, which may be linked to its involvement in protein transport from the Golgi apparatus to the endoplasmic reticulum.Src is activated by various cell surface receptors and phosphorylates numerous cellular proteins (for review, see Refs. 1 and 2). A significant fraction of Src is localized to the perinuclear region and the secretory vesicles in neuronal cells (3,4). It interacts with and phosphorylates proteins such as dynamin and synapsin (5-7) and synaptophysin (8) or cellugyrin (9), suggesting a role in the regulation of membrane traffic. Src and other Src family tyrosine kinases are also localized to the Golgi apparatus (3, 10). On the Golgi apparatus, Src interacts with the ubiquitin ligase Cbl, and Src activation leads to an increased recruitment of Cbl specifically at the Golgi. This indicates that Src-Cbl signaling might occur at this organelle (10). Further support for this proposal stems from recent findings that show that Src is involved in the activation of Ras on the Golgi apparatus following mitogenic stimulation (11). Src is also known to phosphorylate and interact with a Golgi apparatus-specific protein, Golgin 67 (12). However, the function of Src signaling at the Golgi remains unknown. The number of signaling molecules that associate with the Golgi membranes and regulate its function and organization is increasing and is known to include protein kinase D, which regulates Golgi-toplasma membrane trafficking (13) (14), and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) and polo-like kinase, which are essential for the fragmentation of the Golgi at the onset of mitosis (15) (16) (17). It is thus reasonable to propose that Src might also be involved in the structural and functional regulation of the Golgi membranes.A major issue in studies involving Src is the functional redundancy between the Src kinase family, which includes nine members (18). Knock-out studies have revea...

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Immunolocalization of the cellular src protein in interphase and mitotic NIH c-src overexpresser cells.

Cited by 129 publications

References 64 publications

Interactions of TOM1L1 with the Multivesicular Body Sorting Machinery

Interactions of TOM1L1 with the Multivesicular Body Sorting Machinery

Src kinase regulates the integrity and function of the Golgi apparatus via activation of dynamin 2

Src Regulates Golgi Structure and KDEL Receptor-dependent Retrograde Transport to the Endoplasmic Reticulum

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