15-Deoxy-Delta(12,14)-prostaglandin J(2) stabilizes, but functionally inactivates p53 by binding to the cysteine 277 residue.
15-Deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a representative cyclopentenone prostaglandin, has many interesting biological effects. In this study, treatment of human breast cancer cells (MCF-7) with 15d-PGJ(2) led to accumulation of p53 protein. However, the p53 DNA binding and its transcriptional activity were significantly reduced. 15d-PGJ(2) directly modified p53 as verified by reacting recombinant p53 with biotinylated 15d-PGJ(2). 9,10-Dihydro-15-deoxy-Delta(12,14)-prostaglandin J(2) lacking the electrophilic alpha,beta-unsaturated functionality failed to inhibit p53 DNA binding as well as to modify p53. Moreover, by conducting an in vitro [(35)S]-labeled p53 translation assay, we identified cysteine 277 as a putative site of p53 modification by 15d-PGJ(2). The DNA-binding ability of a mutant p53 in which cysteine 277 was substituted by alanine was virtually unaffected by 15d-PGJ(2). Likewise, p53 binding activity of biotinylated 15d-PGJ(2) was abolished in mutant cells. In addition, cells expressing wild-type p53 exhibited p53 protein stability to a greater extent than mutant C277A cells. In conclusion, 15d-PGJ(2) can undergo nucleophilic addition to p53, presumably at the cysteine 277 residue, rendering this tumor suppressor less susceptible to proteasomal degradation.
Kim
D-H
DH
College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
Kim
E-H
EH
Na
H-K
HK
Sun
Y
Y
Surh
Y-J
YJ
D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry; D051546:Cyclooxygenase 2 / Q000502:physiology; D003545:Cysteine / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry; D004247:DNA / Q000378:metabolism; D006801:Humans; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000502:physiology; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000494:pharmacology; D011487:Protein Conformation; D051736:Proto-Oncogene Proteins c-mdm2 / Q000502:physiology; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000502:physiology
C097240:15-deoxy-delta(12,14)-prostaglandin J2; D016159:Tumor Suppressor Protein p53; D004247:DNA; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D003545:Cysteine; D015230:Prostaglandin D2