Small guanosine triphosphatase Rho/Rho-associated kinase as a novel regulator of intracellular redistribution of lysosomes in invasive tumor cells

Yukio Nishimura, Kazuyuki Itoh, Kiyoko Yoshioka, Masayoshi Uehata, Masaru Himeno

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

To investigate the role of RhoA on the intracellular membrane dynamics of lysosomes in rat hepatoma cells (MM1), we analyzed the localization of lysosomal aspartic proteinase cathepsin D by confocal immunofluorescence microscopy in the dominant active RhoA-transfected cells. Here we show that the transfection of the dominant active form of human small guanosine triphosphatase (GTPase) RhoA in MM1 cells, a highly invasive cell line, causes the redistribution and spreading of small punctate structures stained for cathepsin D throughout the cytoplasm. We found that the microtubule organization was markedly different in the two cell lines: uniformly developed and polymerized microtubule filaments were seen in the mock transfectants; however, the dynamic organization of microtubules was less pronounced in the active RhoA transfectants. Furthermore, we found for the first time that a selective inhibitor of Rho-associated kinase (p160ROCK), Y-27632, impeded the subcellular spreading of cathepsin D staining and promoted reclustering of cathepsin D toward the perinuclear region in the active RhoA-transfected cells. To our knowledge, this is the first indication that the RhoA/ROCK-mediated signaling pathway is involved in the intracellular membrane dynamics of lysosomes by regulating the cytoskeletal microtubule organization as well as the actin cytoskeletons.

Original languageEnglish
Pages (from-to)341-351
Number of pages11
JournalCell and tissue research
Volume301
Issue number3
DOIs
Publication statusPublished - Jan 1 2000

Fingerprint

rho-Associated Kinases
Cathepsin D
Guanosine
Lysosomes
Microtubules
Intracellular Membranes
Neoplasms
Aspartic Acid Proteases
Cell Line
Actin Cytoskeleton
Fluorescence Microscopy
Confocal Microscopy
Transfection
Hepatocellular Carcinoma
Cytoplasm
Staining and Labeling

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

Cite this

Small guanosine triphosphatase Rho/Rho-associated kinase as a novel regulator of intracellular redistribution of lysosomes in invasive tumor cells. / Nishimura, Yukio; Itoh, Kazuyuki; Yoshioka, Kiyoko; Uehata, Masayoshi; Himeno, Masaru.

In: Cell and tissue research, Vol. 301, No. 3, 01.01.2000, p. 341-351.

Research output: Contribution to journalArticle

Nishimura, Yukio ; Itoh, Kazuyuki ; Yoshioka, Kiyoko ; Uehata, Masayoshi ; Himeno, Masaru. / Small guanosine triphosphatase Rho/Rho-associated kinase as a novel regulator of intracellular redistribution of lysosomes in invasive tumor cells. In: Cell and tissue research. 2000 ; Vol. 301, No. 3. pp. 341-351.
@article{80d1911c24f64dfaa2dd31d937cf81da,
title = "Small guanosine triphosphatase Rho/Rho-associated kinase as a novel regulator of intracellular redistribution of lysosomes in invasive tumor cells",
abstract = "To investigate the role of RhoA on the intracellular membrane dynamics of lysosomes in rat hepatoma cells (MM1), we analyzed the localization of lysosomal aspartic proteinase cathepsin D by confocal immunofluorescence microscopy in the dominant active RhoA-transfected cells. Here we show that the transfection of the dominant active form of human small guanosine triphosphatase (GTPase) RhoA in MM1 cells, a highly invasive cell line, causes the redistribution and spreading of small punctate structures stained for cathepsin D throughout the cytoplasm. We found that the microtubule organization was markedly different in the two cell lines: uniformly developed and polymerized microtubule filaments were seen in the mock transfectants; however, the dynamic organization of microtubules was less pronounced in the active RhoA transfectants. Furthermore, we found for the first time that a selective inhibitor of Rho-associated kinase (p160ROCK), Y-27632, impeded the subcellular spreading of cathepsin D staining and promoted reclustering of cathepsin D toward the perinuclear region in the active RhoA-transfected cells. To our knowledge, this is the first indication that the RhoA/ROCK-mediated signaling pathway is involved in the intracellular membrane dynamics of lysosomes by regulating the cytoskeletal microtubule organization as well as the actin cytoskeletons.",
author = "Yukio Nishimura and Kazuyuki Itoh and Kiyoko Yoshioka and Masayoshi Uehata and Masaru Himeno",
year = "2000",
month = "1",
day = "1",
doi = "10.1007/s004410000243",
language = "English",
volume = "301",
pages = "341--351",
journal = "Cell and Tissue Research",
issn = "0302-766X",
publisher = "Springer Verlag",
number = "3",

}

TY - JOUR

T1 - Small guanosine triphosphatase Rho/Rho-associated kinase as a novel regulator of intracellular redistribution of lysosomes in invasive tumor cells

AU - Nishimura, Yukio

AU - Itoh, Kazuyuki

AU - Yoshioka, Kiyoko

AU - Uehata, Masayoshi

AU - Himeno, Masaru

PY - 2000/1/1

Y1 - 2000/1/1

N2 - To investigate the role of RhoA on the intracellular membrane dynamics of lysosomes in rat hepatoma cells (MM1), we analyzed the localization of lysosomal aspartic proteinase cathepsin D by confocal immunofluorescence microscopy in the dominant active RhoA-transfected cells. Here we show that the transfection of the dominant active form of human small guanosine triphosphatase (GTPase) RhoA in MM1 cells, a highly invasive cell line, causes the redistribution and spreading of small punctate structures stained for cathepsin D throughout the cytoplasm. We found that the microtubule organization was markedly different in the two cell lines: uniformly developed and polymerized microtubule filaments were seen in the mock transfectants; however, the dynamic organization of microtubules was less pronounced in the active RhoA transfectants. Furthermore, we found for the first time that a selective inhibitor of Rho-associated kinase (p160ROCK), Y-27632, impeded the subcellular spreading of cathepsin D staining and promoted reclustering of cathepsin D toward the perinuclear region in the active RhoA-transfected cells. To our knowledge, this is the first indication that the RhoA/ROCK-mediated signaling pathway is involved in the intracellular membrane dynamics of lysosomes by regulating the cytoskeletal microtubule organization as well as the actin cytoskeletons.

AB - To investigate the role of RhoA on the intracellular membrane dynamics of lysosomes in rat hepatoma cells (MM1), we analyzed the localization of lysosomal aspartic proteinase cathepsin D by confocal immunofluorescence microscopy in the dominant active RhoA-transfected cells. Here we show that the transfection of the dominant active form of human small guanosine triphosphatase (GTPase) RhoA in MM1 cells, a highly invasive cell line, causes the redistribution and spreading of small punctate structures stained for cathepsin D throughout the cytoplasm. We found that the microtubule organization was markedly different in the two cell lines: uniformly developed and polymerized microtubule filaments were seen in the mock transfectants; however, the dynamic organization of microtubules was less pronounced in the active RhoA transfectants. Furthermore, we found for the first time that a selective inhibitor of Rho-associated kinase (p160ROCK), Y-27632, impeded the subcellular spreading of cathepsin D staining and promoted reclustering of cathepsin D toward the perinuclear region in the active RhoA-transfected cells. To our knowledge, this is the first indication that the RhoA/ROCK-mediated signaling pathway is involved in the intracellular membrane dynamics of lysosomes by regulating the cytoskeletal microtubule organization as well as the actin cytoskeletons.

UR - http://www.scopus.com/inward/record.url?scp=0033837823&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033837823&partnerID=8YFLogxK

U2 - 10.1007/s004410000243

DO - 10.1007/s004410000243

M3 - Article

C2 - 10994780

AN - SCOPUS:0033837823

VL - 301

SP - 341

EP - 351

JO - Cell and Tissue Research

JF - Cell and Tissue Research

SN - 0302-766X

IS - 3

ER -