Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the rho family of small G proteins

Isamu Okamoto, Yoshiaki Kawano, Mitsuhiro Matsumoto, Moritaka Suga, Kozo Kaibuchi, Masayuki Ando, Hideyuki Saya

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

CD44 is a cell surface receptor for several extracellular matrix components and is implicated in tumor cell invasion and metastasis. Our previous studies have shown that CD44 expressed in cancer cells is proteolytically cleaved at the extracellular domain through membrane- associated metalloproteases and that CD44 cleavage plays a critical role in CD44-mediated tumor cell migration (Okamoto, I., Kawano, Y., Tsuiki, H., Sasaki, J., Nakao, M., Matsumoto, M., Suga, M., Ando, M., Nakajima, M., and Saya, H. (1999) Oncogene 18, 1435-1446). In the present study, we first demonstrate rapid degradation of the membrane-tethered CD44 cleavage product through intracellular proteolytic pathways, and it occurs only after CD44 extracellular cleavage. To address the mechanisms regulating CD44 cleavage at the extracellular domain, we show that 12-O-tetradecanoylphorbol 13-acetate (TPA) and the calcium ionophore ionomycin rapidly enhance metalloprotease- mediated CD44 cleavage in U251MG cells via protein kinase C-dependent and - independent pathways, respectively, suggesting the existence of multiple distinct pathways for regulation of CD44 cleavage. Concomitant with TPA- induced CD44 cleavage, TPA treatment induces redistribution of CD44 and ERM proteins (ezrin, radixin, and moesin) to newly generated membrane ruffling areas. Treatment with lysophosphatidic acid, which is known to activate the Rho-dependent pathway, inhibits TPA-induced CD44 redistribution and CD44 cleavage. Furthermore, overexpression of Rac dominant active mutants results in the redistribution of CD44 to the Rac-induced ruffling areas and the enhancement of CD44 cleavage. These results suggest that the Rho family proteins play a role in regulation of CD44 distribution and cleavage.

Original languageEnglish
Pages (from-to)25525-25534
Number of pages10
JournalJournal of Biological Chemistry
Volume274
Issue number36
DOIs
Publication statusPublished - Sep 3 1999

Fingerprint

Monomeric GTP-Binding Proteins
Tetradecanoylphorbol Acetate
Protein Kinase C
Acetates
Cells
Metalloproteases
Calcium
Membranes
Tumors
Ionomycin
Calcium Ionophores
Cell Surface Receptors
Neoplasms
Proteins
Oncogenes
Cell Movement
Extracellular Matrix
Degradation
Neoplasm Metastasis

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the rho family of small G proteins. / Okamoto, Isamu; Kawano, Yoshiaki; Matsumoto, Mitsuhiro; Suga, Moritaka; Kaibuchi, Kozo; Ando, Masayuki; Saya, Hideyuki.

In: Journal of Biological Chemistry, Vol. 274, No. 36, 03.09.1999, p. 25525-25534.

Research output: Contribution to journalArticle

Okamoto, Isamu ; Kawano, Yoshiaki ; Matsumoto, Mitsuhiro ; Suga, Moritaka ; Kaibuchi, Kozo ; Ando, Masayuki ; Saya, Hideyuki. / Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the rho family of small G proteins. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 36. pp. 25525-25534.
@article{12ce27b40a114fc9bcd5855fa866d1ca,
title = "Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the rho family of small G proteins",
abstract = "CD44 is a cell surface receptor for several extracellular matrix components and is implicated in tumor cell invasion and metastasis. Our previous studies have shown that CD44 expressed in cancer cells is proteolytically cleaved at the extracellular domain through membrane- associated metalloproteases and that CD44 cleavage plays a critical role in CD44-mediated tumor cell migration (Okamoto, I., Kawano, Y., Tsuiki, H., Sasaki, J., Nakao, M., Matsumoto, M., Suga, M., Ando, M., Nakajima, M., and Saya, H. (1999) Oncogene 18, 1435-1446). In the present study, we first demonstrate rapid degradation of the membrane-tethered CD44 cleavage product through intracellular proteolytic pathways, and it occurs only after CD44 extracellular cleavage. To address the mechanisms regulating CD44 cleavage at the extracellular domain, we show that 12-O-tetradecanoylphorbol 13-acetate (TPA) and the calcium ionophore ionomycin rapidly enhance metalloprotease- mediated CD44 cleavage in U251MG cells via protein kinase C-dependent and - independent pathways, respectively, suggesting the existence of multiple distinct pathways for regulation of CD44 cleavage. Concomitant with TPA- induced CD44 cleavage, TPA treatment induces redistribution of CD44 and ERM proteins (ezrin, radixin, and moesin) to newly generated membrane ruffling areas. Treatment with lysophosphatidic acid, which is known to activate the Rho-dependent pathway, inhibits TPA-induced CD44 redistribution and CD44 cleavage. Furthermore, overexpression of Rac dominant active mutants results in the redistribution of CD44 to the Rac-induced ruffling areas and the enhancement of CD44 cleavage. These results suggest that the Rho family proteins play a role in regulation of CD44 distribution and cleavage.",
author = "Isamu Okamoto and Yoshiaki Kawano and Mitsuhiro Matsumoto and Moritaka Suga and Kozo Kaibuchi and Masayuki Ando and Hideyuki Saya",
year = "1999",
month = "9",
day = "3",
doi = "10.1074/jbc.274.36.25525",
language = "English",
volume = "274",
pages = "25525--25534",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "36",

}

TY - JOUR

T1 - Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the rho family of small G proteins

AU - Okamoto, Isamu

AU - Kawano, Yoshiaki

AU - Matsumoto, Mitsuhiro

AU - Suga, Moritaka

AU - Kaibuchi, Kozo

AU - Ando, Masayuki

AU - Saya, Hideyuki

PY - 1999/9/3

Y1 - 1999/9/3

N2 - CD44 is a cell surface receptor for several extracellular matrix components and is implicated in tumor cell invasion and metastasis. Our previous studies have shown that CD44 expressed in cancer cells is proteolytically cleaved at the extracellular domain through membrane- associated metalloproteases and that CD44 cleavage plays a critical role in CD44-mediated tumor cell migration (Okamoto, I., Kawano, Y., Tsuiki, H., Sasaki, J., Nakao, M., Matsumoto, M., Suga, M., Ando, M., Nakajima, M., and Saya, H. (1999) Oncogene 18, 1435-1446). In the present study, we first demonstrate rapid degradation of the membrane-tethered CD44 cleavage product through intracellular proteolytic pathways, and it occurs only after CD44 extracellular cleavage. To address the mechanisms regulating CD44 cleavage at the extracellular domain, we show that 12-O-tetradecanoylphorbol 13-acetate (TPA) and the calcium ionophore ionomycin rapidly enhance metalloprotease- mediated CD44 cleavage in U251MG cells via protein kinase C-dependent and - independent pathways, respectively, suggesting the existence of multiple distinct pathways for regulation of CD44 cleavage. Concomitant with TPA- induced CD44 cleavage, TPA treatment induces redistribution of CD44 and ERM proteins (ezrin, radixin, and moesin) to newly generated membrane ruffling areas. Treatment with lysophosphatidic acid, which is known to activate the Rho-dependent pathway, inhibits TPA-induced CD44 redistribution and CD44 cleavage. Furthermore, overexpression of Rac dominant active mutants results in the redistribution of CD44 to the Rac-induced ruffling areas and the enhancement of CD44 cleavage. These results suggest that the Rho family proteins play a role in regulation of CD44 distribution and cleavage.

AB - CD44 is a cell surface receptor for several extracellular matrix components and is implicated in tumor cell invasion and metastasis. Our previous studies have shown that CD44 expressed in cancer cells is proteolytically cleaved at the extracellular domain through membrane- associated metalloproteases and that CD44 cleavage plays a critical role in CD44-mediated tumor cell migration (Okamoto, I., Kawano, Y., Tsuiki, H., Sasaki, J., Nakao, M., Matsumoto, M., Suga, M., Ando, M., Nakajima, M., and Saya, H. (1999) Oncogene 18, 1435-1446). In the present study, we first demonstrate rapid degradation of the membrane-tethered CD44 cleavage product through intracellular proteolytic pathways, and it occurs only after CD44 extracellular cleavage. To address the mechanisms regulating CD44 cleavage at the extracellular domain, we show that 12-O-tetradecanoylphorbol 13-acetate (TPA) and the calcium ionophore ionomycin rapidly enhance metalloprotease- mediated CD44 cleavage in U251MG cells via protein kinase C-dependent and - independent pathways, respectively, suggesting the existence of multiple distinct pathways for regulation of CD44 cleavage. Concomitant with TPA- induced CD44 cleavage, TPA treatment induces redistribution of CD44 and ERM proteins (ezrin, radixin, and moesin) to newly generated membrane ruffling areas. Treatment with lysophosphatidic acid, which is known to activate the Rho-dependent pathway, inhibits TPA-induced CD44 redistribution and CD44 cleavage. Furthermore, overexpression of Rac dominant active mutants results in the redistribution of CD44 to the Rac-induced ruffling areas and the enhancement of CD44 cleavage. These results suggest that the Rho family proteins play a role in regulation of CD44 distribution and cleavage.

UR - http://www.scopus.com/inward/record.url?scp=0033520361&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033520361&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.36.25525

DO - 10.1074/jbc.274.36.25525

M3 - Article

C2 - 10464284

AN - SCOPUS:0033520361

VL - 274

SP - 25525

EP - 25534

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 36

ER -