Different behavior of l-afadin and neurabin-II during the formation and destruction of cell-cell adherens junction.
We have recently isolated two novel actin filament-binding proteins, l-afadin and neurabin-II and shown that they are localized at cell-cell adherens junction (AJ) in epithelial cells. We found here that l-afadin, neurabin-II, ZO-1, and E-cadherin showed similar and different behavior during the formation and destruction of cell-cell AJ in MDCK cells. In MDCK cells, the accumulation of both l-afadin and E-cadherin, but not that of ZO-1, changed in parallel depending on Rac small G protein activity. Dissociation of MDCK cells by culturing the cells at 2 microM Ca2+ caused rapid endocytosis of E-cadherin, but not that of l-afadin or ZO-1. Addition of phorbol 12-myristate 13-acetate to these dissociated cells formed a tight junction-like structure where ZO-1 and l-afadin, but not neurabin-II or E-cadherin, accumulated. We furthermore found that, in non-epithelial EL cells, which expressed E-cadherin and attached to each other, l-afadin, neurabin-II, ZO-1 and E-cadherin were all localized at AJ. In cadherin-deficient L cells, I-afadin was mainly localized at cell-cell contact sites, but ZO-1 was mainly localized at the tip area of cell processes. Neurabin-II did not accumulate at the plasma membrane area. Neither l-afadin nor neurabin-II significantly interacted with alpha-, beta-catenin, E-cadherin, ZO-1 or occludin.
Sakisaka
T
T
Department of Molecular Biology and Biochemistry, Osaka University Medical School, Suite, Japan.
Nakanishi
H
H
Takahashi
K
K
Mandai
K
K
Miyahara
M
M
Satoh
A
A
Takaishi
K
K
Takai
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002846:Chromatography, Affinity; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D004705:Endocytosis / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism; D007668:Kidney; D016547:Kinesins; D007739:L Cells; D008565:Membrane Proteins / Q000502:physiology; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism* / Q000502:physiology*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009218:Myosins; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism*; D015534:Trans-Activators; D062826:Zonula Occludens-1 Protein; D051178:alpha Catenin; D051176:beta Catenin; D020744:rac GTP-Binding Proteins; D000099273:Neurabins
D015820:Cadherins; D002118:Calcium; D003598:Cytoskeletal Proteins; D019204:GTP-Binding Proteins; D016547:Kinesins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009218:Myosins; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D013755:Tetradecanoylphorbol Acetate; D015534:Trans-Activators; D062826:Zonula Occludens-1 Protein; D051178:alpha Catenin; D051176:beta Catenin; D020744:rac GTP-Binding Proteins; D000099273:Neurabins; C490657:Afdn protein, mouse; C495267:CTNNB1 protein, mouse; C495272:Ctnna1 protein, mouse; C569043:Tjp1 protein, mouse; C108835:afadin