Ring finger protein 149 is an E3 ubiquitin ligase active on wild-type v-Raf murine sarcoma viral oncogene homolog B1 (BRAF).
Members of the RAF family (ARAF, BRAF, and CRAF/RAF-1) are involved in a variety of cellular activities, including growth, survival, differentiation, and transformation. An oncogene encodes BRAF, the function of which is linked to MEK activation. BRAF is the most effective RAF kinase in terms of induction of MEK/ERK activity. However, the mechanisms involved in BRAF regulation remain unclear. In the present work, we used a tandem affinity purification approach to show that RNF149 (RING finger protein 149) interacts with wild-type BRAF. The latter protein is a RING domain-containing E3 ubiquitin ligase involved in control of gene transcription, translation, cytoskeletal organization, cell adhesion, and epithelial development. We showed that RNF149 bound directly to the C-terminal kinase-containing domain of wild-type BRAF and induced ubiquitination, followed by proteasome-dependent degradation, of the latter protein. Functionally, RNF149 attenuated the increase in cell growth induced by wild-type BRAF. However, RNF149 did not bind to mutant BRAF or induce ubiquitination thereof. Thus, we show that RNF149 is an E3 ubiquitin ligase active on wild-type BRAF.
Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Songpa-gu, Seoul 138-736, Republic of Korea.
Seung-Woo
SW
Hong
Dong-Hoon
DH
Jin
Jae-Sik
JS
Shin
Jai-Hee
JH
Moon
Young-Soon
YS
Na
Kyung-Ah
KA
Jung
Seung-Mi
SM
Kim
Jin Cheon
JC
Kim
Kyu-Pyo
KP
Kim
Yong Sang
YS
Hong
Jae-Lyun
JL
Lee
Eun Kyung
EK
Choi
Jung Shin
JS
Lee
Tae Won
TW
Kim
D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival; D045325:HCT116 Cells; D057809:HEK293 Cells; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D019821:Simvastatin / Q000494:pharmacology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination / Q000187:drug effects
D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D019821:Simvastatin; C575382:RNF149 protein, human; D044767:Ubiquitin-Protein Ligases; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D020928:Mitogen-Activated Protein Kinases; D046988:Proteasome Endopeptidase Complex