Adducin: Ca++-dependent association with sites of cell-cell contact.

Kaiser, Horst; O’Keefe, Edward J.; Bennett, Vann

doi:10.1083/jcb.109.2.557

Cited by 145 publications

(118 citation statements)

References 37 publications

(56 reference statements)

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“…The data base search using MS-Tag, based on the sequence tags and the molecular masses, revealed the protein as mouse ␤-adducin with high relatively. In addition, the apparent molecular mass of ␤-adducin on SDS-PAGE was consistent with that reported previously (26), despite the discrepancy between the apparent (116 kDa) and calculated molecular mass of ␤-adducin (80 kDa). These results strongly suggested PI5 for ␤-adducin.…”

Section: Resultssupporting

confidence: 90%

Interaction of the SH2 Domain of Fyn with a Cytoskeletal Protein, β-Adducin

Shima

Okumura

Takao

et al. 2001

Journal of Biological Chemistry

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Fyn is a Src family tyrosine kinase expressed abundantly in neurons and believed to have specific functions in the brain. To understand the function of Fyn tyrosine kinase, we attempted to identify Fyn Src homology 2 (SH2) domain-binding proteins from a Nonidet P-40-insoluble fraction of the mouse brain. ␤-Adducin, an actin filament-associated cytoskeletal protein, was isolated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry. ␤-Adducin was tyrosine phosphorylated by coexpression with wild type but not with a kinase-negative form of Fyn in COS-7 cells. Cell staining analysis showed that coexpression of ␤-adducin with Fyn induced translocation of ␤-adducin from the cytoplasm to the periphery of the cells where it was colocalized with actin filaments and Fyn. These findings suggest that tyrosine-phosphorylated ␤-adducin associates with the SH2 domain of Fyn and colocalizes under plasma membranes.Neurons in mammalian brain express Src family tyrosine kinases including Src, Fyn, Lyn, and Yes, which have been implicated in neuronal development, neuronal network formation, and synaptic transmission. In contrast to their diverse functions, mice lacking Src, Lyn, or Yes did not show any prominent neurological phenotype, probably because they can compensate each other (1). However, loss of the Fyn gene resulted in several behavioral abnormalities accompanied by changes in the construction of the neuronal network (2-7). We have recently found that Fyn-deficient mice have longer free running periods in the circadian rhythm of locomotive activity and a disorder of neuronal distribution in the suprachiasmatic nucleus, a master circadian oscillator (8). These findings suggest that Fyn might have specific and essential functions in the brain.One possible function of these kinases is to regulate cytoskeletal organization and cell-to-cell contact. Rearrangement of the actin cytoskeletal structure was observed in cells transformed by v-Src (9) and those deficient in C-terminal Src family kinase (CSK), 1 the negative regulator of Src family kinases (10, 11). Cells bearing activated forms of Src family kinases exhibit lower stress fiber formation and weaker cell contacts. This may be partly the result of direct phosphorylation of cytoskeletal proteins at their tyrosine residues by these kinases.Src family kinases form complexes with several cytoskeletal components and signaling molecules at the site of cell contacts through their Src homology 2 (SH2) and Src homology 3 (SH3) domains. When integrins attach to the extracellular matrix, focal adhesion kinase is activated and undergoes autophosphorylation of its Tyr-397 (12), which composes an SH2 domain binding site for Src family tyrosine kinases, leading to phosphorylation at Tyr-925 by Src family kinases (13,14). p130cas is another docking protein that binds to the SH3 and SH2 domains of Src kinases (15,16). This binding facilitates the phosphorylation of p130 cas by Src family kinases (16). It has been shown recently that certain signal transductio...

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Section: Resultssupporting

confidence: 90%

Interaction of the SH2 Domain of Fyn with a Cytoskeletal Protein, β-Adducin

Shima

Okumura

Takao

et al. 2001

Journal of Biological Chemistry

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“…The normal targeting of E-cadherin to cell-cell junctions suggests that E-cadherin sorts to the plasma membrane by a pathway independent from betaospectrin, and it is consistent with observations that E-cadherin is concentrated at cell-cell contact before appearance of spectrin (McNeill et al, 1993). Ankyrin and adducin associate with spectrin and are colocalized with spectrin at sites of cell-cell contact in epithelial cells (Nelson and Veshnock, 1987a;Nelson and Hammerton, 1989;Kaiser et al, 1989). Na÷,K+-ATPase interacts with ankyrin in in vitro assays, and it is colocalized with spectrin and ankyrin in epithelial cells (Nelson and Veshnock, 1987b;Koob et al, 1988;Morrow et al, 1989).…”

Section: Localization Of Actin Ankyrin Adducin Na+k+-atpase and supporting

confidence: 85%

Expression of functional domains of beta G-spectrin disrupts epithelial morphology in cultured cells.

Moorthy

Bennett

1995

The Journal of Cell Biology

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Abstract. Spectrin is a major structural protein associated with the cytoplasmic surface of plasma membranes of many types of cells. To study the functions of spectrin, we transfected Caco-2 intestinal epithelial cells with a plasmid conferring neomycin resistance and encoding either actin-binding or ankyrin-binding domains of beta~-spectrin fused with beta-galactosidase. These polypeptides, in principle, could interfere with the interaction of spectrin with actin or ankyrin, as well as block normal assembly of alpha-and betaspectrin subunits. Cells expressing the fusion proteins represented only a small fraction of neomycin-resistant cells, but they could be detected based on expression of beta-galactosidase. Cells expressing spectrin domains exhibited a progressive decrease in amounts of endogenous betac-spectrin, although alpha-spectrin was still present. Betac-spectrin-deficient cells lost epithelial cell morphology, became multinucleated, and eventually disappeared after 10-14 d in culture. Spectrin-associated membrane proteins, ankyrin and adducin, as well as the Na÷,K÷-ATPase, which binds to ankyrin, exhibited altered distributions in cells transfected with betac-spectrin domains. E-cadherin and F-actin, in contrast to ankyrin, adducin, and the Na÷,K÷-ATPase, were expressed, and they exhibited unaltered distribution in betao-spectrin-deficient cells. Cells transfected with the same plasmid encoding betagalactosidase alone survived in culture as the major population of neomycin-resistant cells, and they exhibited no change in morphology or in the distribution of spectrin-associated membrane proteins. These results establish that betac-spectrin is essential for the normal morphology of epithelial cells, as well as for their maintenance in monolayer culture. S PV.CTRIN is an elongated actin-binding protein that is the principal component of a system of structural proteins associated with the cytoplasmic surface of plasma membranes of most metazoan cells (reviewed by Bennett and Gilligan, 1993). Spectrin is comprised of two subunits, termed alpha and beta, that are aligned side-to-side to form heterodimers, and the dimers are linked head-to-head to form tetramers. Beta subunits contain most of the recognition sites of spectrin for other proteins including ankyrin, protein 4.1 actin, as well as the site for ankyrin-independent association of spectrin with membranes. Beta~-spectrin is the most common type of beta subunit, and it is expressed in most vertebrate tissues (Hu et al., 1992). The structure and function of spectrin has been best characterized in mammalian erythrocytes from both in vivo and in vitro studies (Palek and Lambert, 1990;Delaunay and Dhermy, 1993;Gallagher and Forget, 1993;Bennett and Gilligan, 1993). Erythrocyte spectrin forms a membrane-associated polygonal network by the associations of spectrin with actin illaments and with integral membrane proteins through linkages with ankyrin and protein 4.1. Defects and deficiencies in components of the spectrin-actin network result in abnormally fr...

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“…Loss of ␤-adducin results in increased fragility of erythrocyte membranes (Gilligan et al, 1999;Chen et al, 2007). Adducin is widely expressed in most cell types, including epithelial cells where it is localized at the lateral membrane (Kaiser et al, 1989). Adducin is a target for protein kinase C (PKC) and Rho-kinase and can also be regulated with calcium and calmodulin (Ling et al, 1986;Kuhlman et al, 1996;Matsuoka et al, 1996;Fukata et al, 1999).…”

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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Adducin: Ca++-dependent association with sites of cell-cell contact.

Cited by 145 publications

References 37 publications

Interaction of the SH2 Domain of Fyn with a Cytoskeletal Protein, β-Adducin

Interaction of the SH2 Domain of Fyn with a Cytoskeletal Protein, β-Adducin

Expression of functional domains of beta G-spectrin disrupts epithelial morphology in cultured cells.

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

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