徳島大学 教育・研究者情報データベース(EDB)

1. (英) Nakatsuji Hiroyoshi

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2. 西村 範行

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3. (英) Yamamura Rie

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4. (英) Kanayama Hiro-omi

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5. 佐々木 卓也([徳島大学])

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(英) Involvement of actinin-4 in the recruitment of JRAB/MICAL-L2 to cell-cell junctions and the formation of functional tight junctions.

(英) Tight junctions (TJs) are cell-cell adhesive structures that undergo continuous remodeling. We previously demonstrated that Rab13 and a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) localized at TJs and mediated the endocytic recycling of the integral TJ protein occludin and the formation of functional TJs. Here, we investigated how JRAB/MICAL-L2 was targeted to TJs. Using a series of deletion mutants, we found the plasma membrane (PM)-targeting domain within JRAB/MICAL-L2. We then identified actinin-4, which was originally isolated as an actin-binding protein associated with cell motility and cancer invasion/metastasis, as a binding protein for the PM-targeting domain of JRAB/MICAL-L2, using a yeast two-hybrid system. Actinin-4 was colocalized with JRAB/MICAL-L2 at cell-cell junctions and linked JRAB/MICAL-L2 to F-actin. Although actinin-4 bound to JRAB/MICAL-L2 without Rab13, the actinin-4-JRAB/MICAL-L2 interaction was enhanced by Rab13 activation. Depletion of actinin-4 by using small interfering RNA inhibited the recruitment of occludin to TJs during the Ca(2+) switch. During the epithelial polarization after replating, JRAB/MICAL-L2 was recruited from the cytosol to cell-cell junctions. This JRAB/MICAL-L2 recruitment as well as the formation of functional TJs was delayed in actinin-4-depleted cells. These results indicate that actinin-4 is involved in recruiting JRAB/MICAL-L2 to cell-cell junctions and forming functional TJs.

1. (英) Actinin
2. (英) Actins
3. (英) Animals
4. (英) Base Sequence
5. (英) Binding Sites
6. (英) Calcium
7. (英) Cell Line
8. (英) Cell Polarity
9. (英) Cricetinae
10. (英) Cytoskeletal Proteins
11. (英) Epithelial Cells
12. (英) Genetic Vectors
13. (英) Intercellular Junctions
14. (英) Membrane Proteins
15. (英) Mice
16. (英) Microfilament Proteins
17. (英) Plasmids
18. (英) Protein Binding
19. (英) Protein Structure, Tertiary
20. (英) RNA, Small Interfering
21. (英) Recombinant Proteins
22. (英) Sequence Deletion
23. (英) Tight Junctions
24. (英) Transfection
25. (英) Two-Hybrid System Techniques
26. (英) rab GTP-Binding Proteins

1. (英) Article.Affiliation: Department of Biochemistry, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8503, Japan.

2. (英) Article.PublicationTypeList.PublicationType: Journal Article

3. (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't

4. (英) OtherID: PMC2423145