RESUMO
The phosphatase CDC25A is a key regulator of cell cycle progression by dephosphorylating and activating cyclin-CDK complexes. CDC25A is an unstable protein expressed from G1 until mitosis. CDC25A overexpression, which can be caused by stabilization of the protein, accelerates the G1/S and G2/M transitions, leading to genomic instability and promoting tumorigenesis. Thus, controlling CDC25A protein levels by regulating its stability is a critical mechanism for timing cell cycle progression and to maintain genomic integrity. Herein, we show that CDC25A is phosphorylated on Ser40 throughout the cell cycle and that this phosphorylation is established during the progression from G1 to S phase. We demonstrate that CyclinD-CDK4/CDK6 complexes mediate the phosphorylation of CDC25A on Ser40 during G1 and that these complexes directly phosphorylate this residue in vitro. Importantly, we also find that CyclinD1-CDK4 decreases CDC25A stability in a ßTrCP-dependent manner and that Ser40 and Ser88 phosphorylations contribute to this regulation. Thus our results identify cyclinD-CDK4/6 complexes as novel regulators of CDC25A stability during G1 phase, generating a negative feedback loop allowing control of the G1/S transition.
Assuntos
Ciclina D/metabolismo , Quinase 4 Dependente de Ciclina/metabolismo , Quinase 6 Dependente de Ciclina/metabolismo , Fosfatases cdc25/metabolismo , Pontos de Checagem do Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Ciclina D/genética , Quinase 4 Dependente de Ciclina/genética , Quinase 6 Dependente de Ciclina/genética , Fase G1/fisiologia , Células HEK293 , Humanos , Fosforilação , Estabilidade Proteica , Fase S/fisiologia , Transfecção , Fosfatases cdc25/genéticaRESUMO
We report on the use of a thulium-fiber-pumped holmium-based emitter in a coherent differential absorption lidar (CDIAL) experiment for high time and space resolution of CO(2) absorption field in the atmosphere. The 2-µm high-power dual-wavelength single-mode Q-switched Ho:YLF oscillator delivers 10-mJ pulses with a duration of 40 ns at 2 kHz. Both short pulse duration and high repetition rate were chosen to increase the DIAL precision and time and space resolution in coherent detection. The CDIAL provides 150-m range and 15-min time-resolved CO(2) absorption coefficient with a calculated instrumental error of 0.5% at 500 m and less than 2% at 1 km. Dry-air CO(2) mixing ratio estimates from the DIAL system are compared with simultaneous in situ gas analyzer measurements during a 20-h-long experiment.
