Rho regulates association of both the ERM family and vinculin with the plasma membrane in MDCK cells

被引:66
作者
Kotani, H [1 ]
Takaishi, K [1 ]
Sasaki, T [1 ]
Takai, Y [1 ]
机构
[1] OSAKA UNIV,SCH MED,DEPT MOL BIOL & BIOCHEM,SUITA,OSAKA 565,JAPAN
关键词
Rho; ERM family; vinculin; peripheral bundle; cortical bundle; focal adhesion;
D O I
10.1038/sj.onc.1200998
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Rho small G protein regulates various actin-dependent cell functions. As to the functioning sites of Rho, Rho regulates formation of stress fibers and focal adhesions in many types of cultured cells, whereas we have shown that the association sites of actin filaments with the plasma membrane controlled by the ERM (Ezrin, Radixin, Moesin) family are the functioning sites of Rho in MDCK cells stably expressing myc-RhoA. We have investigated here the effect of microinjection of Rho GDI, a negative regulator of Rho which inhibits activation of Rho, C3, an exoenzyme of Clostridium botulinum which ADP-ribosylates Rho and inhibits its functions, or guanosine 5'-(3-O-thio) triphosphate-bound active form of Rho on the intracellular localization of both the ERM family and vinculin, which is one of the structural proteins of focal adhesions, in wild type MDCK cells. The ERM family was preferentially localized at peripheral bundles of actin filaments which are localized at the outer edge of colonies of the cells, microvilli and low Gaze-induced cortical bundles of actin filaments in wild type MDCK cells. Microinjection of Rho GDI or C3 inhibited the localization of the ERM family at both the peripheral bundles and the low Ca2+-induced cortical bundles. On the other hand, vinculin was localized at both focal adhesions and basal edges of the colonies of the cells, and microinjection of Rho GDI or C3 inhibited the localization of vinculin at both of these sites. These results indicate that activation of Rho is necessary for the association of both the ERM family and vinculin with the plasma membrane in wild type MDCK cells. Microinjection of the guanosine 5'-(3-O-thio) triphosphate-bound form of Rho induced an increase in the localization of vinculin at focal adhesions, but did not induce an increase in the localization of the ERM family at the plasma membrane, indicating that activation of Rho itself is sufficient only for the association of vinculin with the plasma membrane at focal adhesions.
引用
收藏
页码:1705 / 1713
页数:9
相关论文
共 47 条
  • [1] BOTULINUM ADP-RIBOSYLTRANSFERASE C-3 - PURIFICATION OF THE ENZYME AND CHARACTERIZATION OF THE ADP-RIBOSYLATION REACTION IN PLATELET MEMBRANES
    AKTORIES, K
    ROSENER, S
    BLASCHKE, U
    CHHATWAL, GS
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 172 (02): : 445 - 450
  • [2] Identification of a putative target for Rho as the serine-threonine kinase protein kinase N
    Amano, M
    Mukai, H
    Ono, Y
    Chihara, K
    Matsui, T
    Hamajima, Y
    Okawa, K
    Iwamatsu, A
    Kaibuchi, K
    [J]. SCIENCE, 1996, 271 (5249) : 648 - 650
  • [3] PURIFICATION OF THE 22 KDA PROTEIN SUBSTRATE OF BOTULINUM ADP-RIBOSYLTRANSFERASE C-3 FROM PORCINE BRAIN CYTOSOL AND ITS CHARACTERIZATION AS A GTP-BINDING PROTEIN HIGHLY HOMOLOGOUS TO THE RHO GENE-PRODUCT
    BRAUN, U
    HABERMANN, B
    JUST, I
    AKTORIES, K
    VANDEKERCKHOVE, J
    [J]. FEBS LETTERS, 1989, 243 (01) : 70 - 76
  • [5] FOCAL ADHESIONS - TRANSMEMBRANE JUNCTIONS BETWEEN THE EXTRACELLULAR-MATRIX AND THE CYTOSKELETON
    BURRIDGE, K
    FATH, K
    KELLY, T
    NUCKOLLS, G
    TURNER, C
    [J]. ANNUAL REVIEW OF CELL BIOLOGY, 1988, 4 : 487 - 525
  • [6] Rho1p, a yeast protein at the interface between cell polarization and morphogenesis
    Drgonova, J
    Drgon, T
    Tanaka, K
    Kollar, R
    Chen, GC
    Ford, RA
    Chan, CSM
    Takai, Y
    Cabib, E
    [J]. SCIENCE, 1996, 272 (5259) : 277 - 279
  • [7] FUKUMOTO Y, 1990, ONCOGENE, V5, P1321
  • [8] VINCULIN, AN INTRACELLULAR PROTEIN LOCALIZED AT SPECIALIZED SITES WHERE MICROFILAMENT BUNDLES TERMINATE AT CELL-MEMBRANES
    GEIGER, B
    TOKUYASU, KT
    DUTTON, AH
    SINGER, SJ
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (07): : 4127 - 4131
  • [9] SMALL GTP-BINDING PROTEINS AND THE REGULATION OF THE ACTIN CYTOSKELETON
    HALL, A
    [J]. ANNUAL REVIEW OF CELL BIOLOGY, 1994, 10 : 31 - 54
  • [10] Regulation mechanism of ERM (ezrin/radixin/moesin) protein/plasma membrane association: Possible involvement of phosphatidylinositol turnover and Rho-dependent signaling pathway
    Hirao, M
    Sato, N
    Kondo, T
    Yonemura, S
    Monden, M
    Sasaki, T
    Takai, Y
    Tsukita, S
    Tsukita, S
    [J]. JOURNAL OF CELL BIOLOGY, 1996, 135 (01) : 37 - 51