Rab11-FIP2 influences multiple components of the endosomal system in polarized MDCK cells

Ducharme, Nicole A.; Ham, Amy-Joan L.; Lapierre, Lynne A.; Goldenring, James R.

doi:10.4161/cl.1.2.15289

Cited by 23 publications

(19 citation statements)

References 31 publications

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“…Previous studies have shown that a tripartite targeting complex consisting of MyoVb, Rab11a and Rab11-FIP2 mediates the surface expression of many apical proteins (Chu et al, 2009;Ducharme et al, 2011;Hales et al, 2002;Lindsay and McCaffrey, 2002;Nedvetsky et al, 2007). Loss of MyoVb function causes mislocalization of ABC transporters involved in the maintenance of apical polarity in hepatocytes to Rab11a-rich subapical endosomes (Wakabayashi et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Myosin Vb mediates Cu+ export in polarized hepatocytes

Gupta

Schell

Bhattacharjee

et al. 2016

Journal of Cell Science

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The cellular machinery responsible for Cu + -stimulated delivery of the Wilson-disease-associated protein ATP7B to the apical domain of hepatocytes is poorly understood. We demonstrate that myosin Vb regulates the Cu + -stimulated delivery of ATP7B to the apical domain of polarized hepatic cells, and that disruption of the ATP7B-myosin Vb interaction reduces the apical surface expression of ATP7B. Overexpression of the myosin Vb tail, which competes for binding of subapical cargos to myosin Vb bound to subapical actin, disrupted the surface expression of ATP7B, leading to reduced cellular Cu + export. The myosin-Vb-dependent targeting step occurred in parallel with hepatocyte-like polarity. If the myosin Vb tail was expressed acutely in cells just prior to the establishment of polarity, it appeared as part of an intracellular apical compartment, centered on γ-tubulin. ATP7B became selectively arrested in this compartment at high [Cu + ] in the presence of myosin Vb tail, suggesting that these compartments are precursors of donor-acceptor transfer stations for apically targeted cargos of myosin Vb. Our data suggest that reduced hepatic Cu + clearance in idiopathic non-Wilsonian types of disease might be associated with the loss of function of myosin Vb.

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Section: Introductionmentioning

confidence: 99%

Myosin Vb mediates Cu+ export in polarized hepatocytes

Gupta

Schell

Bhattacharjee

et al. 2016

Journal of Cell Science

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“…25 Through sequence analysis of the Rab11-FIPs, FIP1B and FIP1C were identified to contain the same MARK2 phosphorylation consensus sequence as FIP2 (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

“…22,23 Rab11-FIP2 interacting with Rab11 and Myosin Vb is important for plasma membrane recycling, regulating trafficking at multiple steps during transcytosis and apical recycling. 20,24,25 Phosphorylation of Rab11-FIP2 by MARK2 at serine 227 is upregulated early in junction re-assembly following readdition of extracellular calcium after an overnight incubation in low calcium medium (calcium switch), and overexpression of a phospho-mimetic (S227E) form of Rab11-FIP2 in MDCK cells causes a multi-lumen phenotype in 3-dimenstional cyst culture. 26 Interestingly, pS227-Rab11-FIP2…”

Section: Introductionmentioning

confidence: 99%

Rab11-FIP1 phosphorylation by MARK2 regulates polarity in MDCK cells

et al. 2017

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MARK2/Par1b/EMK1, a serine/threonine kinase, is required for correct apical/basolateral membrane polarization in epithelial cells. However, the specific substrates mediating MARK2 action are less well understood. We have now found that MARK2 phosphorylates Rab11-FIP1B/C at serine 234 in a consensus site similar to that previously identified in Rab11-FIP2. In MDCK cells undergoing repolarization after a calcium switch, antibodies specific for pS234-Rab11-FIP1 or pS227-Rab11-FIP2 demonstrate that the spatial and temporal activation of Rab11-FIP1 phosphorylation is distinct from that for Rab11-FIP2. Phosphorylation of Rab11-FIP1 persists through calcium switch and remains high after polarity has been reestablished whereas FIP2 phosphorylation is highest early in reestablishment of polarity but significantly reduced once polarity has been re-established. MARK2 colocalized with FIP1B/C/D and p(S234)-FIP1 in vivo. Overexpression of GFP-Rab11-FIP1C wildtype or nonphosphorylatable GFP-Rab11-FIP1C(S234A) induced two significant phenotypes following calcium switch. Overexpression of FIP1C wildtype and FIP1C(S234A) caused a psuedo-stratification of cells in early time points following calcium switch. At later time points most prominently observed in cells expressing FIP1C(S234A) a significant lateral lumen phenotype was observed, where F-actin-rich lateral lumens appeared demarcated by a ring of ZO1 and also containing ezrin, syntaxin 3 and podocalyxin. In contrast, p120 and E-Cadherin were excluded from the new apical surface at the lateral lumens and now localized to the new lateral surface oriented toward the media. GFP-FIP1C(S234A) localized to membranes deep to the lateral lumens, and immunostaining demonstrated the reorientation of the centrosome and the Golgi apparatus toward the lateral lumen. These results suggest that both Rab11-FIP1B/C and Rab11-FIP2 serve as critical substrates mediating aspects of MARK2 regulation of epithelial polarity.

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“…Two mutants in Rab11-FIP2, Rab11-FIP2(SARG) and Rab11-FIP2(129-512), inhibit trafficking through the apical recycling system in MDCK cells [12, 32]. Rab11-FIP2(SARG) appears to act at a more distal point in the apical recycling system.…”

Section: Discussionmentioning

confidence: 99%

“…Cherry-Rab11a was described in Baetz et al[16] Cherry-Rab5a was described in Ducharme et al[32] GFP-FIP2 was originally described in Hales et al[19] From this construct, Rab11-FIP2 was subcloned into Cherry-N2 to create Rab11-FIP2-Cherry. Rab11-FIP1A and Rab11-FIP1A(T197A) were subcloned into pET30a for protein purification.…”

Section: Methodsmentioning

confidence: 99%

Rab11-FIP1A regulates early trafficking into the recycling endosomes

Schafer¹,

McRae²,

Manning³

et al. 2016

Experimental Cell Research

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The Rab11 family of small GTPases, along with the Rab11-family interacting proteins (Rab11-FIPs), are critical regulators of intracellular vesicle trafficking and recycling. We have identified a point mutation of Threonine-197 site to an Alanine in Rab11-FIP1A, which causes a dramatic dominant negative phenotype when expressed in HeLa cells. The normally perinuclear distribution of GFP-Rab11-FIP1A was condensed into a membranous cisternum with almost no GFP-Rab11-FIP1A(T197A) remaining outside of this central locus. Also, this condensed GFP-FIP1A(T197A) altered the distribution of proteins in the Rab11a recycling pathway including endogenous Rab11a, Rab11-FIP1C, and transferrin receptor (CD71). Furthermore, this condensed GFP-FIP1A(T197A)-containing structure exhibited little movement in live HeLa cells. Expression of GFP-FIP1A(T197A) caused a strong blockade of transferrin recycling. Treatment of cells expressing GFP-FIP1A(T197A) with nocodazole did not disperse the Rab11a-containing recycling system. We also found that Rab5 and EEA1 were accumulated in membranes by GFP-Rab11-FIP1A but Rab4 was unaffected, suggesting that a direct pathway may exist from early endosomes into the Rab11a-containing recycling system. Our study of a potent inhibitory trafficking mutation in Rab11-FIP1A shows that Rab11-FIP1A associates with and regulates trafficking at an early step in the process of membrane recycling.

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Rab11-FIP2 influences multiple components of the endosomal system in polarized MDCK cells

Cited by 23 publications

References 31 publications

Myosin Vb mediates Cu+ export in polarized hepatocytes

Myosin Vb mediates Cu+ export in polarized hepatocytes

Rab11-FIP1 phosphorylation by MARK2 regulates polarity in MDCK cells

Rab11-FIP1A regulates early trafficking into the recycling endosomes

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