Lateral Membrane Biogenesis in Human Bronchial Epithelial Cells Requires 190-kDa Ankyrin-G

Kizhatil, Krishnakumar; Bennett, Vann

doi:10.1074/jbc.m314296200

Cited by 89 publications

(111 citation statements)

References 46 publications

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“…An unresolved question for the future is whether ankyrin-G acts solely as a localized scaffolding protein that stabilizes Na v 1.5 at specialized sites once it is delivered to the plasma membrane, or whether ankyrin-G also has a role in sorting and trafficking of these channels. The recent finding that ankyrin-G is required for biogenesis of the lateral membrane domain of epithelial cells (22) suggests functions for ankyrin-G that go beyond simple scaffolding.…”

Section: Discussionmentioning

confidence: 99%

Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes

Mohler

Rivolta

Napolitano

et al. 2004

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

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336

357

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We identify a human mutation (E1053K) in the ankyrin-binding motif of Na v1.5 that is associated with Brugada syndrome, a fatal cardiac arrhythmia caused by altered function of Na v1.5. The E1053K mutation abolishes binding of Na v1.5 to ankyrin-G, and also prevents accumulation of Na v1.5 at cell surface sites in ventricular cardiomyocytes. Ankyrin-G and Nav1.5 are both localized at intercalated disc and T-tubule membranes in cardiomyocytes, and Na v1.5 coimmunoprecipitates with 190-kDa ankyrin-G from detergent-soluble lysates from rat heart. These data suggest that Na v1.5 associates with ankyrin-G and that ankyrin-G is required for Na v1.5 localization at excitable membranes in cardiomyocytes. Together with previous work in neurons, these results in cardiomyocytes suggest that ankyrin-G participates in a common pathway for localization of voltage-gated Na v channels at sites of function in multiple excitable cell types.arrhythmia ͉ SCN5A ͉ targeting ͉ trafficking ͉ sudden cardiac death

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Section: Discussionmentioning

confidence: 99%

Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes

Mohler

Rivolta

Napolitano

et al. 2004

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

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336

357

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“…Loss of ankyrin-G or ␤2-spectrin abolishes de novo formation of lateral membranes during mitosis (Kizhatil and Bennett, 2004;Kizhatil et al, 2007a), whereas loss of adducin has no effect ( Figure 3B). Moreover, cells depleted of either ankyrin-G or ␤2-spectrin accumulate E-cadherin in the transGolgi (Kizhatil et al, 2007b).…”

Section: Adducin Is Necessary To Stabilize Preformed Lateral Membranesmentioning

confidence: 99%

“…Moreover, resident lateral membrane proteins including the Na/K ATPase and E-cadherin either bind directly or coimmunoprecipitate with ankyrin and spectrin (Nelson and Veshnock, 1987;Nelson and Hammerton, 1989;Nelson et al, 1990;Morrow et al, 1989;Davis and Bennett, 1990). Recently, ␤2-spectrin and ankyrin have been demonstrated to be required for both stability and biogenesis of the lateral membrane of cultured epithelial cells (Kizhatil and Bennett, 2004;Kizhatil et al, 2007a). Furthermore, E-cadherin has been shown to bind directly to ankyrin-G and to require ankyrin-G and ␤2-spectrin for transport to the lateral membrane (Kizhatil et al, 2007b).…”

Section: Introductionmentioning

confidence: 99%

Adducin Promotes Micrometer-Scale Organization of β2-Spectrin in Lateral Membranes of Bronchial Epithelial Cells

Abdi

Bennett

2008

MBoC

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Adducin promotes assembly of spectrin-actin complexes, and is a target for regulation by calmodulin, protein kinase C, and rho kinase. We demonstrate here that adducin is required to stabilize preformed lateral membranes of human bronchial epithelial (HBE) cells through interaction with ␤2-spectrin. We use a Tet-on regulated inducible small interfering RNA (siRNA) system to deplete ␣-adducin from confluent HBE cells. Depletion of ␣-adducin resulted in increased detergent solubility of spectrin after normal membrane biogenesis during mitosis. Conversely, depletion of ␤2-spectrin resulted in loss of adducin from the lateral membrane. siRNA-resistant ␣-adducin prevented loss of lateral membrane, but only if ␣-adducin retained the MARCKS domain that mediates spectrin-actin interactions. Phosphomimetic versions of adducin with S/D substitutions at protein kinase C phosphorylation sites in the MARCKS domain were not active in rescue. We find that adducin modulates long-range organization of the lateral membrane based on several criteria. First, the lateral membrane of adducin-depleted cells exhibited reduced height, increased curvature, and expansion into the basal surface. Moreover, E-cadherin-GFP, which normally is restricted in lateral mobility, rapidly diffuses over distances up to 10 m. We conclude that adducin acting through spectrin provides a novel mechanism to regulate global properties of the lateral membrane of bronchial epithelial cells.

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“…Targeted knock-out of ankyrin-G in the mouse cerebellum results in loss of clustering of the voltage-gated sodium channel Na v 1.6 and L1 CAMs neurofascin and NrCAM at Purkinje neuron initial segments (3,4). Knock-down of ankyrin-G in cultured epithelial cells by small interfering RNA results in loss of the lateral membrane domain (5). Loss of ankyrin-B in mice results in deficiency of Na,K-ATPase, Na/Ca exchanger, and inositol 1,4,5-trisphosphate (InsP 3 ) 1 receptor located in a specialized microdomain of T-tubules in adult cardiomyocytes (6).…”

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confidence: 99%

Ankyrin-B Targets β2-Spectrin to an Intracellular Compartment in Neonatal Cardiomyocytes

Mohler

Yoon

Bennett

2004

Journal of Biological Chemistry

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107

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Ankyrin-B is a spectrin-binding protein that is required for localization of inositol 1,4,5-trisphosphate receptor and ryanodine receptor in neonatal cardiomyocytes. This work addresses the interaction between ankyrin-B and ␤ 2 -spectrin in these cells. Ankyrin-B and ␤ 2 -spectrin are colocalized in an intracellular striated compartment overlying the M-line and distinct from Ttubules, sarcoplasmic reticulum, Golgi, endoplasmic reticulum, lysosomes, and endosomes. ␤ 2 -Spectrin is absent in ankyrin-B-null cardiomyocytes and is restored to a normal striated pattern by rescue with green fluorescent protein-220-kDa ankyrin-B. We identified two mutants (A1000P and DAR976AAA) located in the ZU5 domain which eliminate spectrin binding activity of ankyrin-B. Ankyrin-B mutants lacking spectrin binding activity are normally targeted but do not reestablish ␤ 2 -spectrin in ankyrin-B ؉/؊ cardiomyocytes. However, both mutant forms of ankyrin-B are still capable of restoring inositol 1,4,5-trisphosphate receptor localization and normal contraction frequency of cardiomyocytes. Therefore, direct binding of ␤ 2 -spectrin to ankyrin-B is required for the normal targeting of ␤ 2 -spectrin in neonatal cardiomyocytes. In contrast, ankyrin-B localization and function are independent of ␤ 2 -spectrin. In summary, this work demonstrates that interaction between members of the ankyrin and ␤-spectrin families previously established in erythrocytes and axon initial segments also occurs in neonatal cardiomyocytes with ankyrin-B and ␤ 2 -spectrin. This work also establishes a functional hierarchy in which ankyrin-B determines the localization of ␤ 2 -spectrin and operates independently of ␤ 2 -spectrin in its role in organizing membrane-spanning proteins.

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Lateral Membrane Biogenesis in Human Bronchial Epithelial Cells Requires 190-kDa Ankyrin-G

Cited by 89 publications

References 46 publications

Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes

Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes

Adducin Promotes Micrometer-Scale Organization of β2-Spectrin in Lateral Membranes of Bronchial Epithelial Cells

Ankyrin-B Targets β2-Spectrin to an Intracellular Compartment in Neonatal Cardiomyocytes

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Lateral Membrane Biogenesis in Human Bronchial Epithelial Cells Requires 190-kDa Ankyrin-G

Cited by 89 publications

References 46 publications

Nav1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Nav1.5 on the surface of cardiomyocytes

Nav1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Nav1.5 on the surface of cardiomyocytes

Adducin Promotes Micrometer-Scale Organization of β2-Spectrin in Lateral Membranes of Bronchial Epithelial Cells

Ankyrin-B Targets β2-Spectrin to an Intracellular Compartment in Neonatal Cardiomyocytes

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Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes

Na_v1.5 E1053K mutation causing Brugada syndrome blocks binding to ankyrin-G and expression of Na_v1.5 on the surface of cardiomyocytes