Complex regulation of cell-cycle inhibitors by Fbxw7 in mouse embryonic fibroblasts.
The F-box protein Fbxw7 (also known as Fbw7, SEL-10, hCdc4 or hAgo) mediates the ubiquitylation and thereby contributes to the degradation of proteins that positively regulate cell cycle. Conditional ablation of Fbxw7 in mouse embryonic fibroblasts (MEFs) induces cell-cycle arrest accompanied by abnormal accumulation of the intracellular domain of Notch1 (NICD1) and c-Myc. However, the molecular mechanisms by which the accumulation of NICD1 and c-Myc induces cell-cycle arrest have remained unclear. We have now examined the expression of cell-cycle inhibitors in Fbxw7-deficient MEFs and found that the abundance of p27(Kip1) and p57(Kip2) is paradoxically decreased. This phenomenon appears to be attributable to the accumulation of NICD1, given that it was recapitulated by overexpression of NICD1 and blocked by ablation of RBP-J. Conversely, the expression of p16(Ink4a) and p19(ARF) was increased in an NICD1-independent manner in Fbxw7-null MEFs. The increased expression of p19(ARF) was recapitulated by overexpression of c-Myc and abolished by ablation of c-Myc, suggesting that the accumulation of c-Myc is primarily responsible for that of p19(ARF). In contrast, the upregulation of p16(Ink4a) appeared to be independent of c-Myc. These results indicate that cell-cycle inhibitors undergo complex regulation by the Fbxw7-mediated proteolytic system.
Masuda
K
K
Department of Developmental Genetics, Center for Translational and Advanced Animal Research, Graduate School of Medicine, Tohoku University, Aoba-ku, Sendai 980-8575, Japan.
Ishikawa
Y
Y
Onoyama
I
I
Unno
M
M
de Alborán
I M
IM
Nakayama
K I
KI
Nakayama
K
K
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000187:drug effects / Q000235:genetics; D050761:Cyclin-Dependent Kinase Inhibitor p57 / Q000187:drug effects / Q000235:genetics; D017931:DNA Primers; D015536:Down-Regulation / Q000187:drug effects; D044783:F-Box Proteins / Q000235:genetics / Q000494:pharmacology*; D000073979:F-Box-WD Repeat-Containing Protein 7; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D006706:Homeostasis / Q000187:drug effects; D051379:Mice; D018345:Mice, Knockout; D016271:Proto-Oncogene Proteins c-myc / Q000494:pharmacology; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics / Q000494:pharmacology*; D054875:Ubiquitination
C493961:Cdkn1b protein, mouse; D050761:Cyclin-Dependent Kinase Inhibitor p57; D017931:DNA Primers; D044783:F-Box Proteins; D000073979:F-Box-WD Repeat-Containing Protein 7; C513272:Fbxw7 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D050760:Cyclin-Dependent Kinase Inhibitor p27; D044767:Ubiquitin-Protein Ligases