Src-dependent phosphorylation of caveolin-1 Tyr-14 promotes swelling and release of caveolae

Zimnicka, Adriana M.; Husain, Yawer; Shajahan, Ayesha N.; Sverdlov, Maria; Chaga, Oleg Y.; Chen, Zhenlong; Tóth, Péter T.; Klomp, Jennifer E.; Karginov, Andrei V.; Tiruppathì, Chinnaswamy; Malik, Asrar B.; Minshall, Richard D.

doi:10.1091/mbc.e15-11-0756

Cited by 105 publications

(109 citation statements)

References 69 publications

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“…These data indicate that Tyr-940 is a key residue involved in the intramolecular interaction, and consistently it was reported that Tyr-940 of DENND3 is phosphorylated in rat brain (22). Because of the aromatic ring of tyrosine, there is no natural amino acid substitute that creates an ideal phosphomimetic mutant, but many examples do exist in which mutation of Tyr to Asp or Glu has been used successfully to mimic phosphorylated tyrosine (23)(24)(25)(26)(27). Theoretically, if the effect of the phosphorylation is due primarily to the negative charge of the phospho group, Asp or Glu should mimic phosphorylated tyrosine.…”

Section: Identification Of Tyr-940 As a Key Residue Involved In The Isupporting

confidence: 67%

Regulation of DENND3, the exchange factor for the small GTPase Rab12 through an intramolecular interaction

McPherson

2017

Journal of Biological Chemistry

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Edited by Thomas SöllnerThe Rab family of small GTPases functions in multiple aspects of cellular membrane trafficking. Proteins bearing a differentially expressed in normal and neoplastic cells (DENN) domain have emerged as the largest family of Rab-activating guanine nucleotide exchange factors (GEFs). Rab12 functions in the initiation of starvation-induced autophagy, and our previous work revealed that its activator, DENN domain-containing protein 3 (DENND3), is phosphorylated and activated upon starvation. However, how the GEF activity of DENND3 toward Rab12 is regulated at the molecular level is still not understood. Here, we combine size-exclusion chromatography, Förster resonance energy transfer, pulldown, and in vitro GEF assays to demonstrate that regulation of GEF activity is achieved through an intramolecular interaction that is controlled by a key residue in DENND3, tyrosine 940. Our study sheds light on the regulation of Rab12 activation and lays the groundwork for characterizing the regulation of other DENN domain-containing proteins.The Rab family of small GTPases functions in all aspects of cellular membrane trafficking ranging from vesicle budding to transport along the cytoskeleton and fusion with acceptor membranes. Rabs switch between an inactive GDP-bound form and an active GTP-bound form that interacts with effectors to mediate trafficking functions. Guanine nucleotide exchange factors (GEFs) 3 activate Rabs by facilitating the exchange of GDP for GTP (1). There are at least four families of GEFs for Rabs: TRAPP, Rabin8/Sec2, Vps9 domain-containing proteins, and DENN domain-containing proteins (2). With 26 distinct members, DENN domain-containing proteins are the largest family of Rab GEFs (3-8). They are involved in diverse biological functions (4), and mutations in several DENN domain-bearing proteins are linked to human diseases, including the tumor suppressor folliculin associated with Birt-HoggDubé syndrome (9), C9orf72 linked to familial frontotemporal dementia and amyotrophic lateral sclerosis (10, 11), and DENND1B associated with childhood asthma (12).We recently demonstrated that DENN domain-containing protein 3 (DENND3) functions in starvation-induced macroautophagy (13,14). Macroautophagy, which we will hereafter refer to as autophagy, is a conserved cellular process in which various physiological signals, including nutrient starvation, lead to the recruitment of organelles or cytosolic proteins into double membrane vesicles called autophagosomes. The autophagosomes eventually fuse with lysosomes for degradation of the internalized cargo. The resulting building blocks, such as amino acids, are then released back to the cytosol for reuse, helping cells survive starvation. Unc-51-like kinase (ULK) is the most upstream kinase for autophagy initiation (15). Upon starvation, ULK phosphorylates DENND3 at Ser-554 and Ser-572, recruiting the adapter protein 14-3-3, and this process leads to up-regulation of DENND3 GEF activity toward its substrate Rab12. Rab12 on recycling endosomes (1...

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Section: Identification Of Tyr-940 As a Key Residue Involved In The Isupporting

confidence: 67%

Regulation of DENND3, the exchange factor for the small GTPase Rab12 through an intramolecular interaction

McPherson

2017

Journal of Biological Chemistry

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“…In general, Src-mediated Cav1 phosphorylation on Tyr14 has been demonstrated to be essential for Cav1 signaling and Cav1 endocytosis because phosphorylation of Cav1 leads to separation of neighboring negatively-charged N-terminal phospho-tyrosine residues, promoting swelling of caveolae followed by their release from the plasma membrane4748. Hereby Src has also been shown to play an important role in PCa development and progression because Src can signal through focal adhesion kinase (FAK) in response to integrin activation, which has been implicated in many aspects of tumor biology, such as cell proliferation, metastasis and angiogenesis4950.…”

Section: Discussionmentioning

confidence: 99%

Progression-related loss of stromal Caveolin 1 levels fosters the growth of human PC3 xenografts and mediates radiation resistance

Panic

Ketteler

Reis

et al. 2017

Sci Rep

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Despite good treatment results in localized prostate tumors, advanced disease stages usually have a pronounced resistance to chemotherapy and radiotherapy. The membrane protein caveolin-1 (Cav1) functions here as an important oncogene. Therefore we examined the impact of stromal Cav1 expression for tumor growth and sensitivity to ionizing radiation (IR). Silencing of Cav1 expression in PC3 cells resulted in increased tumor growth and a reduced growth delay after IR when compared to tumors generated by Cav1-expressing PC3 cells. The increased radiation resistance was associated with increasing amounts of reactive tumor stroma and a Cav1 re-expression in the malignant epithelial cells. Mimicking the human situation these results were confirmed using co-implantation of Cav1-silenced PC3 cells with Cav1-silenced or Cav1-expressing fibroblasts. Immunohistochemically analysis of irradiated tumors as well as human prostate tissue specimen confirmed that alterations in stromal-epithelial Cav1 expressions were accompanied by a more reactive Cav1-reduced tumor stroma after radiation and within advanced prostate cancer tissues which potentially mediates the resistance to radiation treatment. Conclusively, the radiation response of human prostate tumors is critically regulated by Cav1 expression in stromal fibroblasts. Loss of stromal Cav1 expression in advanced tumor stages may thus contribute to resistance of these tumors to radiotherapy.

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“…eNOS is activated by phosphorylation at multiple sites, and lowered p-eNOS levels indicate a decreased ability to vasodilate and an increased propensity for vessel wall hyperplasia [35,36]. Cav-1 is a scaffolding protein essential in the cellular transport of macromolecules, and its phosphorylation leads to the destabilization of this process [37]. A decrease in the phosphorylation of Cav-1 thus indicates a decreased ability of EC to mediate growth, contributing to vessel wall thickening.…”

Section: Discussionmentioning

confidence: 99%

Perturbations in Endothelial Dysfunction-Associated Pathways in the Nitrofen-Induced Congenital Diaphragmatic Hernia Model

Zhaorigetu

Bair²,

Lu³

et al. 2017

J Vasc Res

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Although it is well known that nitrofen induces congenital diaphragmatic hernia (CDH), including CDH-associated lung hypoplasia and pulmonary hypertension (PH) in rodents, the mechanism of pathogenesis remains largely unclear. It has been reported that pulmonary artery (PA) endothelial cell (EC) dysfunction contributes to the development of PH in CDH. Thus, we hypothesized that there is significant alteration of endothelial dysfunction-associated proteins in nitrofen-induced CDH PAs. Pregnant SD rats received either nitrofen or olive oil on gestational day 9.5. The newborn rats were sacrificed and divided into a CDH (n = 81) and a control (n = 23) group. After PA isolation, the expression of PA endothelial dysfunction-associated proteins was assessed on Western blot and immunostaining. We demonstrate that the expression of C-reactive protein and endothelin-1 and its receptors, ET_A and ET_B, were significantly increased in the CDH PAs. Levels of phosphorylated myosin light chain were significantly elevated, but those of phosphorylated endothelial nitric oxide synthase, caveolin-1, and mechanistic target of rapamycin were significantly decreased in the CDH PAs. In this work, we elucidate alterations in the expression of endothelial dysfunction-associated proteins specific to nitrofen-induced CDHrodent PAs, thereby advancing our understanding of the critical role of endothelial dysfunction-associated pathways in the pathogenesis of nitrofen-induced CDH.

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Src-dependent phosphorylation of caveolin-1 Tyr-14 promotes swelling and release of caveolae

Cited by 105 publications

References 69 publications

Regulation of DENND3, the exchange factor for the small GTPase Rab12 through an intramolecular interaction

Regulation of DENND3, the exchange factor for the small GTPase Rab12 through an intramolecular interaction

Progression-related loss of stromal Caveolin 1 levels fosters the growth of human PC3 xenografts and mediates radiation resistance

Perturbations in Endothelial Dysfunction-Associated Pathways in the Nitrofen-Induced Congenital Diaphragmatic Hernia Model

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