Protection of cisplatin cytotoxicity by an inactive cyclin-dependent kinase.
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.
Hodeify
Rawad
R
Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.
Megyesi
Judit
J
Tarcsafalvi
Adel
A
Safirstein
Robert L
RL
Price
Peter M
PM
D021581:Active Transport, Cell Nucleus; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D001665:Binding Sites; D053148:Caspase 3 / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology; D002478:Cells, Cultured; D002945:Cisplatin / Q000633:toxicity*; D019925:Cyclin A / Q000235:genetics / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism*; D045304:Cytochromes c / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D019610:Cytoprotection; D004706:Endodeoxyribonucleases / Q000378:metabolism; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection
D000970:Antineoplastic Agents; D019925:Cyclin A; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate; D045304:Cytochromes c; C495894:CDK2 protein, human; C495893:Cdk2 protein, mouse; D051357:Cyclin-Dependent Kinase 2; D004706:Endodeoxyribonucleases; C052391:endonuclease G; C505409:Casp3 protein, mouse; D053148:Caspase 3; D002945:Cisplatin
Biochem Biophys Res Commun. 1997 Jan 3;230(1):49-53
9020058
J Exp Med. 1999 Mar 15;189(6):957-68
10075979
J Am Soc Nephrol. 2006 Sep;17(9):2434-42
16914540
J Biol Chem. 2001 Jun 22;276(25):22404-9
11304535
Curr Biol. 2003 Jan 21;13(2):R71-3
12546810
Trends Cell Biol. 2003 Feb;13(2):65-70
12559756
Cell Death Differ. 2004 Dec;11(12):1277-86
15297883
Oncogene. 2008 Oct 27;27(50):6452-61
18955972
Drugs. 2000;59 Suppl 4:9-17; discussion 37-8
10864226
EMBO J. 1998 Dec 15;17(24):7209-18
9857178
Circ Res. 2001 Mar 2;88(4):408-14
11230108
Toxicol Sci. 2001 Oct;63(2):196-207
11568363
Cancer Lett. 2005 Jan 20;217(2):129-38
15617830
Nature. 1970 Aug 15;227(5259):680-5
5432063
Am J Physiol. 1995 Aug;269(2 Pt 1):C323-33
7653514
Cell. 1996 Jul 12;86(1):147-57
8689682
Biochem Biophys Res Commun. 1994 Jun 30;201(3):1072-8
8024548
EMBO J. 2003 Sep 1;22(17):4385-99
12941691
Am J Physiol Renal Physiol. 2008 Jul;295(1):F44-52
18400869
Oncogene. 2002 Mar 7;21(11):1750-8
11896606
Cancer Cell. 2004 Feb;5(2):163-75
14998492
Science. 1993 Dec 24;262(5142):2050-4
8266103
Oncogene. 2006 Jul 6;25(29):4056-66
16491117
Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3754-8
8170983
J Biol Chem. 2003 Mar 14;278(11):9100-6
12509415
Cell Death Differ. 2002 Oct;9(10):1031-42
12232790
Mol Biol Cell. 2002 Mar;13(3):1030-45
11907280
Mol Cell. 1998 Mar;1(4):553-63
9660939
J Biol Chem. 2002 Oct 11;277(41):38476-85
12176996
Semin Nephrol. 2003 Sep;23(5):416-24
13680530
Nature. 1999 Feb 4;397(6718):441-6
9989411
J Clin Invest. 2002 Sep;110(6):835-42
12235115
Am J Physiol Renal Physiol. 2004 Feb;286(2):F378-84
12965891
J Cell Sci. 1993 Oct;106 ( Pt 2):535-44
8282760
Nature. 2001 Jul 5;412(6842):95-9
11452314
Nat Biotechnol. 2004 Dec;22(12):1567-72
15558047
Kidney Int. 2009 Sep;76(6):604-13
19536080
Cell. 1993 Nov 19;75(4):817-25
8242752
J Hepatol. 2000 Aug;33(2):266-74
10952244
Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2509-14
9482916
Am J Physiol Renal Physiol. 2005 Sep;289(3):F514-20
15840769
Am J Physiol Renal Physiol. 2007 Jul;293(1):F52-9
17459956
J Mol Cell Cardiol. 2008 Nov;45(5):610-6
18692063
Science. 2002 Nov 22;298(5598):1587-92
12446902
Histochem Cell Biol. 1999 May;111(5):359-65
10403114
J Clin Invest. 1998 Feb 15;101(4):777-82
9466972
J Am Soc Nephrol. 2005 Jul;16(7):1985-92
15901768