Rac1 regulates heat shock responses by reorganization of vimentin filaments: Identification using MALDI-TOF MS

S. Y. Lee, E. J. Song, H. J. Kim, H. J. Kang, Jae-Hong Kim, K. J. Lee

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Rac1 has been implicated in a wide variety of biological processes, including actin remodeling and various signaling cascades. Here we have examined whether Rac1 might be involved in heat shock-induced cell signaling. We found that Rat2 stable cells expressing a dominant negative Rac1 mutant, RacN17 (Rat2-RacN17), were significantly more tolerant to heat shock than control Rat2 cells, and simultaneously inhibited the activation of SAPK/JNK by heat shock compared to control Rat2 cells. However, no discernible effect was observed in typical heat shock responses including total protein synthesis and heat shock protein synthesis. To identify the proteins involved in this difference, we separated the proteins of both Rat2 and Rat2-RacN17 cell lines after heat shock using two-dimensional gel electrophoresis and identified the differentially expressed proteins by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) after in-gel trypsin digestion. Differentially expressed proteins between two cell lines were identified as vimentin. Rat2-RacN17 cells showed significant changes in vimentin as well as marked changes in vimentin reorganization by heat shock. The vimentin changes were identified as N-terminal head domain cleavage. These results suggest that Rac1 plays a pivotal role in the heat shock-induced signaling cascade by modifying intermediate vimentin filaments.

Original languageEnglish
Pages (from-to)1093-1102
Number of pages10
JournalCell Death and Differentiation
Volume8
Issue number11
DOIs
Publication statusPublished - 2001 Nov 26
Externally publishedYes

Fingerprint

Heat-Shock Response
Vimentin
Shock
Mass Spectrometry
Lasers
Hot Temperature
Proteins
Biological Phenomena
Cell Line
Intermediate Filaments
Electrophoresis, Gel, Two-Dimensional
Heat-Shock Proteins
Trypsin
Actins
Digestion
Gels
Head

Keywords

  • Heat shock
  • MALDI-TOF MS
  • Rac1
  • SAPK/JNK activation
  • Thermotolerance
  • Vimentin

ASJC Scopus subject areas

  • Cell Biology

Cite this

Rac1 regulates heat shock responses by reorganization of vimentin filaments : Identification using MALDI-TOF MS. / Lee, S. Y.; Song, E. J.; Kim, H. J.; Kang, H. J.; Kim, Jae-Hong; Lee, K. J.

In: Cell Death and Differentiation, Vol. 8, No. 11, 26.11.2001, p. 1093-1102.

Research output: Contribution to journalArticle

Lee, S. Y. ; Song, E. J. ; Kim, H. J. ; Kang, H. J. ; Kim, Jae-Hong ; Lee, K. J. / Rac1 regulates heat shock responses by reorganization of vimentin filaments : Identification using MALDI-TOF MS. In: Cell Death and Differentiation. 2001 ; Vol. 8, No. 11. pp. 1093-1102.
@article{d5e63cb97b58437697e428d2198230a8,
title = "Rac1 regulates heat shock responses by reorganization of vimentin filaments: Identification using MALDI-TOF MS",
abstract = "Rac1 has been implicated in a wide variety of biological processes, including actin remodeling and various signaling cascades. Here we have examined whether Rac1 might be involved in heat shock-induced cell signaling. We found that Rat2 stable cells expressing a dominant negative Rac1 mutant, RacN17 (Rat2-RacN17), were significantly more tolerant to heat shock than control Rat2 cells, and simultaneously inhibited the activation of SAPK/JNK by heat shock compared to control Rat2 cells. However, no discernible effect was observed in typical heat shock responses including total protein synthesis and heat shock protein synthesis. To identify the proteins involved in this difference, we separated the proteins of both Rat2 and Rat2-RacN17 cell lines after heat shock using two-dimensional gel electrophoresis and identified the differentially expressed proteins by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) after in-gel trypsin digestion. Differentially expressed proteins between two cell lines were identified as vimentin. Rat2-RacN17 cells showed significant changes in vimentin as well as marked changes in vimentin reorganization by heat shock. The vimentin changes were identified as N-terminal head domain cleavage. These results suggest that Rac1 plays a pivotal role in the heat shock-induced signaling cascade by modifying intermediate vimentin filaments.",
keywords = "Heat shock, MALDI-TOF MS, Rac1, SAPK/JNK activation, Thermotolerance, Vimentin",
author = "Lee, {S. Y.} and Song, {E. J.} and Kim, {H. J.} and Kang, {H. J.} and Jae-Hong Kim and Lee, {K. J.}",
year = "2001",
month = "11",
day = "26",
doi = "10.1038/sj.cdd.4400923",
language = "English",
volume = "8",
pages = "1093--1102",
journal = "Cell Death and Differentiation",
issn = "1350-9047",
publisher = "Nature Publishing Group",
number = "11",

}

TY - JOUR

T1 - Rac1 regulates heat shock responses by reorganization of vimentin filaments

T2 - Identification using MALDI-TOF MS

AU - Lee, S. Y.

AU - Song, E. J.

AU - Kim, H. J.

AU - Kang, H. J.

AU - Kim, Jae-Hong

AU - Lee, K. J.

PY - 2001/11/26

Y1 - 2001/11/26

N2 - Rac1 has been implicated in a wide variety of biological processes, including actin remodeling and various signaling cascades. Here we have examined whether Rac1 might be involved in heat shock-induced cell signaling. We found that Rat2 stable cells expressing a dominant negative Rac1 mutant, RacN17 (Rat2-RacN17), were significantly more tolerant to heat shock than control Rat2 cells, and simultaneously inhibited the activation of SAPK/JNK by heat shock compared to control Rat2 cells. However, no discernible effect was observed in typical heat shock responses including total protein synthesis and heat shock protein synthesis. To identify the proteins involved in this difference, we separated the proteins of both Rat2 and Rat2-RacN17 cell lines after heat shock using two-dimensional gel electrophoresis and identified the differentially expressed proteins by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) after in-gel trypsin digestion. Differentially expressed proteins between two cell lines were identified as vimentin. Rat2-RacN17 cells showed significant changes in vimentin as well as marked changes in vimentin reorganization by heat shock. The vimentin changes were identified as N-terminal head domain cleavage. These results suggest that Rac1 plays a pivotal role in the heat shock-induced signaling cascade by modifying intermediate vimentin filaments.

AB - Rac1 has been implicated in a wide variety of biological processes, including actin remodeling and various signaling cascades. Here we have examined whether Rac1 might be involved in heat shock-induced cell signaling. We found that Rat2 stable cells expressing a dominant negative Rac1 mutant, RacN17 (Rat2-RacN17), were significantly more tolerant to heat shock than control Rat2 cells, and simultaneously inhibited the activation of SAPK/JNK by heat shock compared to control Rat2 cells. However, no discernible effect was observed in typical heat shock responses including total protein synthesis and heat shock protein synthesis. To identify the proteins involved in this difference, we separated the proteins of both Rat2 and Rat2-RacN17 cell lines after heat shock using two-dimensional gel electrophoresis and identified the differentially expressed proteins by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) after in-gel trypsin digestion. Differentially expressed proteins between two cell lines were identified as vimentin. Rat2-RacN17 cells showed significant changes in vimentin as well as marked changes in vimentin reorganization by heat shock. The vimentin changes were identified as N-terminal head domain cleavage. These results suggest that Rac1 plays a pivotal role in the heat shock-induced signaling cascade by modifying intermediate vimentin filaments.

KW - Heat shock

KW - MALDI-TOF MS

KW - Rac1

KW - SAPK/JNK activation

KW - Thermotolerance

KW - Vimentin

UR - http://www.scopus.com/inward/record.url?scp=0035166170&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035166170&partnerID=8YFLogxK

U2 - 10.1038/sj.cdd.4400923

DO - 10.1038/sj.cdd.4400923

M3 - Article

C2 - 11687887

AN - SCOPUS:0035166170

VL - 8

SP - 1093

EP - 1102

JO - Cell Death and Differentiation

JF - Cell Death and Differentiation

SN - 1350-9047

IS - 11

ER -