Phosphorylation of p53 by IkappaB kinase 2 promotes its degradation by beta-TrCP.
Functional inactivation of p53 and constitutive activation of the NF-kappaB pathway has been associated with several human cancers. In this study, we show that IkappaB kinase 2 (IKK2/IKKbeta), which is critical for NF-kappaB activation, also phosphorylates p53. Phosphorylation of p53 at serines 362 and 366 by IKK2 leads to its recruitment to and ubiquitination by beta-TrCP1. Degradation of ubiquitinated p53 is independent of Mdm2, because it occurs in both wild-type and Mdm2(-/-) cells. SiRNA-mediated reduction in the levels of beta-TrCP1 and other members of the SCF(beta-TrCP1)E3 ubiquitin ligase complex or overexpression of a dominant negative form of beta-TrCP1 enhances p53 stability. Substitutions at Ser-362 and 366 of p53 by alanines (p53 AA) result in reduced phosphorylation of p53 by IKK2, decreased association with beta-TrCP1, and thus increased stability of p53 and expression of p53 target genes such as p21, altering the G1 phase of the cell cycle. Our results identify IKK2 and beta-TrCP1 as novel regulators of the p53 pathway and suggest that blocking of IKK2 and beta-TrCP1 could be a means of regulating p53 stability and thereby modulating its biological activity.
Xia
Yifeng
Y
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Padre
Roanna C
RC
De Mendoza
Tatiana Hurtado
TH
Bottero
Virginie
V
Tergaonkar
Vinay B
VB
Verma
Inder M
IM
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D006868:Hydrolysis; D051550:I-kappa B Kinase / Q000378:metabolism*; D051379:Mice; D010449:Peptide Mapping; D010766:Phosphorylation; D011485:Protein Binding; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D054875:Ubiquitination; D044844:beta-Transducin Repeat-Containing Proteins / Q000378:metabolism*
D017931:DNA Primers; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D044844:beta-Transducin Repeat-Containing Proteins; D051550:I-kappa B Kinase