Thapsigargin induces phosphorylation of the 27-kDa heat shock protein in human keratinocytes

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

In the human keratinocyte line HaCaT, the non-phosphorylated 27-kDa heat shock protein (HSP27) isoform A (pI 6.5) is constitutively expressed. Application of thapsigargin, which inhibits Ca2+-ATPase in the endoplasmic reticulum, results in the rapid formation of the phosphorylated HSP27 isoform B (pI 6.0) and reduction of HSP27 A without affecting the synthesis of HSP27. The thapsigargin-dependent HSP27 isoform change is similar to that induced by 43°C heat shock, but different from that induced by arsenite, where the biphosphorylated isoform HSP27 C (pI 5.7) is observed. The receptor agonist bradykinin, which increases intracellular Ca2+ (Ca(i)) level, shows no effect on the distribution of HSP27 isoforms. The responses of HSP27 isoforms to thapsigargin are independent of Ca(i) concentration in HaCaT cells. These observations suggest that the thapsigargin-induced change in HSP27 isoforms is dependent on the depletion of internal Ca2+ stores rather than on the increase in Ca(i) concentration. The thapsigargin-induced change in HSP27 isoforms is reduced by the tyrosine kinase inhibitor, genistein, but not the protein kinase C inhibitor, H-7. We propose that the modulation of HSP27 phosphorylation status by Ca(i) homeostasis may be mechanistically linked to control of keratinocyte growth and differentiation and responses of keratinocytes to extracellular stresses.

Original languageEnglish (US)
Pages (from-to)749-754
Number of pages6
JournalJournal of Investigative Dermatology
Volume107
Issue number5
StatePublished - 1996

Fingerprint

HSP27 Heat-Shock Proteins
Phosphorylation
Thapsigargin
Heat-Shock Proteins
Keratinocytes
Protein Isoforms
Bradykinin Receptors
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
Calcium-Transporting ATPases
Protein C Inhibitor
Genistein
Bradykinin
Protein Kinase Inhibitors
Endoplasmic Reticulum
Protein-Tyrosine Kinases
Protein Kinase C
Shock
Homeostasis
Hot Temperature
Modulation

Keywords

  • HaCaT keratinocytes
  • HSP27 isoforms
  • Intracellular calcium
  • Stress protein

ASJC Scopus subject areas

  • Dermatology

Cite this

Thapsigargin induces phosphorylation of the 27-kDa heat shock protein in human keratinocytes. / Shi, Biao; Isseroff, Roslyn Rivkah.

In: Journal of Investigative Dermatology, Vol. 107, No. 5, 1996, p. 749-754.

Research output: Contribution to journalArticle

@article{ab0563422bf04839b8248429aa1a349b,
title = "Thapsigargin induces phosphorylation of the 27-kDa heat shock protein in human keratinocytes",
abstract = "In the human keratinocyte line HaCaT, the non-phosphorylated 27-kDa heat shock protein (HSP27) isoform A (pI 6.5) is constitutively expressed. Application of thapsigargin, which inhibits Ca2+-ATPase in the endoplasmic reticulum, results in the rapid formation of the phosphorylated HSP27 isoform B (pI 6.0) and reduction of HSP27 A without affecting the synthesis of HSP27. The thapsigargin-dependent HSP27 isoform change is similar to that induced by 43°C heat shock, but different from that induced by arsenite, where the biphosphorylated isoform HSP27 C (pI 5.7) is observed. The receptor agonist bradykinin, which increases intracellular Ca2+ (Ca(i)) level, shows no effect on the distribution of HSP27 isoforms. The responses of HSP27 isoforms to thapsigargin are independent of Ca(i) concentration in HaCaT cells. These observations suggest that the thapsigargin-induced change in HSP27 isoforms is dependent on the depletion of internal Ca2+ stores rather than on the increase in Ca(i) concentration. The thapsigargin-induced change in HSP27 isoforms is reduced by the tyrosine kinase inhibitor, genistein, but not the protein kinase C inhibitor, H-7. We propose that the modulation of HSP27 phosphorylation status by Ca(i) homeostasis may be mechanistically linked to control of keratinocyte growth and differentiation and responses of keratinocytes to extracellular stresses.",
keywords = "HaCaT keratinocytes, HSP27 isoforms, Intracellular calcium, Stress protein",
author = "Biao Shi and Isseroff, {Roslyn Rivkah}",
year = "1996",
language = "English (US)",
volume = "107",
pages = "749--754",
journal = "Journal of Investigative Dermatology",
issn = "0022-202X",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - Thapsigargin induces phosphorylation of the 27-kDa heat shock protein in human keratinocytes

AU - Shi, Biao

AU - Isseroff, Roslyn Rivkah

PY - 1996

Y1 - 1996

N2 - In the human keratinocyte line HaCaT, the non-phosphorylated 27-kDa heat shock protein (HSP27) isoform A (pI 6.5) is constitutively expressed. Application of thapsigargin, which inhibits Ca2+-ATPase in the endoplasmic reticulum, results in the rapid formation of the phosphorylated HSP27 isoform B (pI 6.0) and reduction of HSP27 A without affecting the synthesis of HSP27. The thapsigargin-dependent HSP27 isoform change is similar to that induced by 43°C heat shock, but different from that induced by arsenite, where the biphosphorylated isoform HSP27 C (pI 5.7) is observed. The receptor agonist bradykinin, which increases intracellular Ca2+ (Ca(i)) level, shows no effect on the distribution of HSP27 isoforms. The responses of HSP27 isoforms to thapsigargin are independent of Ca(i) concentration in HaCaT cells. These observations suggest that the thapsigargin-induced change in HSP27 isoforms is dependent on the depletion of internal Ca2+ stores rather than on the increase in Ca(i) concentration. The thapsigargin-induced change in HSP27 isoforms is reduced by the tyrosine kinase inhibitor, genistein, but not the protein kinase C inhibitor, H-7. We propose that the modulation of HSP27 phosphorylation status by Ca(i) homeostasis may be mechanistically linked to control of keratinocyte growth and differentiation and responses of keratinocytes to extracellular stresses.

AB - In the human keratinocyte line HaCaT, the non-phosphorylated 27-kDa heat shock protein (HSP27) isoform A (pI 6.5) is constitutively expressed. Application of thapsigargin, which inhibits Ca2+-ATPase in the endoplasmic reticulum, results in the rapid formation of the phosphorylated HSP27 isoform B (pI 6.0) and reduction of HSP27 A without affecting the synthesis of HSP27. The thapsigargin-dependent HSP27 isoform change is similar to that induced by 43°C heat shock, but different from that induced by arsenite, where the biphosphorylated isoform HSP27 C (pI 5.7) is observed. The receptor agonist bradykinin, which increases intracellular Ca2+ (Ca(i)) level, shows no effect on the distribution of HSP27 isoforms. The responses of HSP27 isoforms to thapsigargin are independent of Ca(i) concentration in HaCaT cells. These observations suggest that the thapsigargin-induced change in HSP27 isoforms is dependent on the depletion of internal Ca2+ stores rather than on the increase in Ca(i) concentration. The thapsigargin-induced change in HSP27 isoforms is reduced by the tyrosine kinase inhibitor, genistein, but not the protein kinase C inhibitor, H-7. We propose that the modulation of HSP27 phosphorylation status by Ca(i) homeostasis may be mechanistically linked to control of keratinocyte growth and differentiation and responses of keratinocytes to extracellular stresses.

KW - HaCaT keratinocytes

KW - HSP27 isoforms

KW - Intracellular calcium

KW - Stress protein

UR - http://www.scopus.com/inward/record.url?scp=0029848314&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029848314&partnerID=8YFLogxK

M3 - Article

VL - 107

SP - 749

EP - 754

JO - Journal of Investigative Dermatology

JF - Journal of Investigative Dermatology

SN - 0022-202X

IS - 5

ER -