Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase

Rawad Hodeify, Judit Megyesi, Adel Tarcsafalvi, Robert L. Safirstein, Peter M. Price

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Cisplatin cytotoxicity is dependent on cyclin-dependent kinase 2 (Cdk2) activity in vivo and in vitro. A Cdk2 mutant (Cdk2-F80G) was designed in which the ATP-binding pocket was altered. When expressed in mouse kidney cells, this protein was kinase inactive, did not inhibit endogenous Cdk2, but protected from cisplatin. The mutant was localized in the cytoplasm, but when coexpressed with cyclin A, it was activated, localized to the nucleus, and no longer protected from cisplatin cytotoxicity. Cells exposed to cisplatin in the presence of the activated mutant had an apoptotic phenotype, and endonuclease G was released from mitochondria similar to that mediated by endogenous Cdk2. But unlike apoptosis mediated by wild-type Cdk2, cisplatin exposure of cells expressing the activated mutant did not cause cytochrome c release or significant caspase-3 activation. We conclude that cisplatin likely activates both caspase-dependent and -independent cell death, and Cdk2 is required for both pathways. The mutant-inactive Cdk2 protected from both death pathways, but after activation by excess cyclin A, caspase-independent cell death predominated.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Physiology
Volume299
Issue number1
DOIs
Publication statusPublished - 1 Jul 2010
Externally publishedYes

Fingerprint

Cyclin-Dependent Kinase 2
Cyclin-Dependent Kinases
Cisplatin
Cyclin A
Caspases
Cell Death
Cytochromes c
Caspase 3
Protein Kinases
Mitochondria
Cytoplasm
Adenosine Triphosphate
Apoptosis
Phenotype
Kidney

Keywords

  • Caspase-3
  • Cell death
  • Cyclin A

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase. / Hodeify, Rawad; Megyesi, Judit; Tarcsafalvi, Adel; Safirstein, Robert L.; Price, Peter M.

In: American Journal of Physiology - Renal Physiology, Vol. 299, No. 1, 01.07.2010.

Research output: Contribution to journalArticle

Hodeify, Rawad ; Megyesi, Judit ; Tarcsafalvi, Adel ; Safirstein, Robert L. ; Price, Peter M. / Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase. In: American Journal of Physiology - Renal Physiology. 2010 ; Vol. 299, No. 1.
@article{c76533a56e4f4bae9f1863bbdfd752bf,
title = "Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase",
abstract = "Cisplatin cytotoxicity is dependent on cyclin-dependent kinase 2 (Cdk2) activity in vivo and in vitro. A Cdk2 mutant (Cdk2-F80G) was designed in which the ATP-binding pocket was altered. When expressed in mouse kidney cells, this protein was kinase inactive, did not inhibit endogenous Cdk2, but protected from cisplatin. The mutant was localized in the cytoplasm, but when coexpressed with cyclin A, it was activated, localized to the nucleus, and no longer protected from cisplatin cytotoxicity. Cells exposed to cisplatin in the presence of the activated mutant had an apoptotic phenotype, and endonuclease G was released from mitochondria similar to that mediated by endogenous Cdk2. But unlike apoptosis mediated by wild-type Cdk2, cisplatin exposure of cells expressing the activated mutant did not cause cytochrome c release or significant caspase-3 activation. We conclude that cisplatin likely activates both caspase-dependent and -independent cell death, and Cdk2 is required for both pathways. The mutant-inactive Cdk2 protected from both death pathways, but after activation by excess cyclin A, caspase-independent cell death predominated.",
keywords = "Caspase-3, Cell death, Cyclin A",
author = "Rawad Hodeify and Judit Megyesi and Adel Tarcsafalvi and Safirstein, {Robert L.} and Price, {Peter M.}",
year = "2010",
month = "7",
day = "1",
doi = "10.1152/ajprenal.00151.2010",
language = "English",
volume = "299",
journal = "American Journal of Physiology - Renal Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "1",

}

TY - JOUR

T1 - Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase

AU - Hodeify, Rawad

AU - Megyesi, Judit

AU - Tarcsafalvi, Adel

AU - Safirstein, Robert L.

AU - Price, Peter M.

PY - 2010/7/1

Y1 - 2010/7/1

N2 - Cisplatin cytotoxicity is dependent on cyclin-dependent kinase 2 (Cdk2) activity in vivo and in vitro. A Cdk2 mutant (Cdk2-F80G) was designed in which the ATP-binding pocket was altered. When expressed in mouse kidney cells, this protein was kinase inactive, did not inhibit endogenous Cdk2, but protected from cisplatin. The mutant was localized in the cytoplasm, but when coexpressed with cyclin A, it was activated, localized to the nucleus, and no longer protected from cisplatin cytotoxicity. Cells exposed to cisplatin in the presence of the activated mutant had an apoptotic phenotype, and endonuclease G was released from mitochondria similar to that mediated by endogenous Cdk2. But unlike apoptosis mediated by wild-type Cdk2, cisplatin exposure of cells expressing the activated mutant did not cause cytochrome c release or significant caspase-3 activation. We conclude that cisplatin likely activates both caspase-dependent and -independent cell death, and Cdk2 is required for both pathways. The mutant-inactive Cdk2 protected from both death pathways, but after activation by excess cyclin A, caspase-independent cell death predominated.

AB - Cisplatin cytotoxicity is dependent on cyclin-dependent kinase 2 (Cdk2) activity in vivo and in vitro. A Cdk2 mutant (Cdk2-F80G) was designed in which the ATP-binding pocket was altered. When expressed in mouse kidney cells, this protein was kinase inactive, did not inhibit endogenous Cdk2, but protected from cisplatin. The mutant was localized in the cytoplasm, but when coexpressed with cyclin A, it was activated, localized to the nucleus, and no longer protected from cisplatin cytotoxicity. Cells exposed to cisplatin in the presence of the activated mutant had an apoptotic phenotype, and endonuclease G was released from mitochondria similar to that mediated by endogenous Cdk2. But unlike apoptosis mediated by wild-type Cdk2, cisplatin exposure of cells expressing the activated mutant did not cause cytochrome c release or significant caspase-3 activation. We conclude that cisplatin likely activates both caspase-dependent and -independent cell death, and Cdk2 is required for both pathways. The mutant-inactive Cdk2 protected from both death pathways, but after activation by excess cyclin A, caspase-independent cell death predominated.

KW - Caspase-3

KW - Cell death

KW - Cyclin A

UR - http://www.scopus.com/inward/record.url?scp=77953835356&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77953835356&partnerID=8YFLogxK

U2 - 10.1152/ajprenal.00151.2010

DO - 10.1152/ajprenal.00151.2010

M3 - Article

VL - 299

JO - American Journal of Physiology - Renal Physiology

JF - American Journal of Physiology - Renal Physiology

SN - 1931-857X

IS - 1

ER -