MEK2 regulates ribonucleotide reductase activity through functional interaction with ribonucleotide reductase small subunit p53R2.
The p53R2 protein, a newly identified member of the ribonucleotide reductase family that provides nucleotides for DNA damage repair, is directly regulated by p53. We show that p53R2 is also regulated by a MEK2 (ERK kinase 2/MAP kinase kinase 2)-dependent pathway. Increased MEK1/2 phosphorylation by serum stimulation coincided with an increase in the RNR activity in U2OS and H1299 cells. The inhibition of MEK2 activity, either by treatment with a MEK inhibitor or by transfection with MEK2 siRNA, dramatically decreased the serum-stimulated RNR activity. Moreover, p53R2 siRNA, but not R2 siRNA, significantly inhibits serum-stimulated RNR activity, indicating that p53R2 is specifically regulated by a MEK2-dependent pathway. Co-immunoprecipitation analyses revealed that the MEK2 segment comprising amino acids 65-171 is critical for p53R2-MEK2 interaction, and the binding domain of MEK2 is required for MEK2-mediated increased RNR activity. Phosphorylation of MEK1/2 was greatly augmented by ionizing radiation, and RNR activity was concurrently increased. Ionizing radiation-induced RNR activity was markedly attenuated by transfection of MEK2 or p53R2 siRNA, but not R2 siRNA. These data show that MEK2 is an endogenous regulator of p53R2 and suggest that MEK2 may associate with p53R2 and upregulate its activity.
DNA Damage Response Network Center, Chosun University School of Medicine, Gwangju, South Korea.
Chunmei
C
Piao
Cha-Kyung
CK
Youn
Min
M
Jin
Sang Pil
SP
Yoon
In-Youb
IY
Chang
Jung Hee
JH
Lee
Ho Jin
HJ
You
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D004260:DNA Repair / Q000235:genetics*; D005720:Gamma Rays; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D047468:Immunoprecipitation; D048370:MAP Kinase Kinase 2 / Q000378:metabolism* / Q000502:physiology; D010766:Phosphorylation; D034622:RNA Interference; D012264:Ribonucleotide Reductases / Q000378:metabolism*; D012588:Scintillation Counting
D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; C405947:RRM2B protein, human; D012264:Ribonucleotide Reductases; C482128:MAP2K2 protein, human; D048370:MAP Kinase Kinase 2