Nitric oxide activation of Erk1/2 regulates the stability and translation of mRNA transcripts containing CU-rich elements

Nitric oxide (NO center dot) can stabilize mRNA by activating p38 mitogen-activated protein kinase (MAPK). Here, transcript stabilization by NO center dot was investigated in human THP-1 cells using microarrays. After LPS pre-stimulation, cells were treated with actinomycin D and then exposed to NO center dot without or with the p38 MAPK inhibitor SB202190 (SB). The decay of 220 mRNAs was affected; most were stabilized by NO% Unexpectedly, SB often enhanced rather than antagonized transcript stability. NO center dot activated p38 MAPK and Erk1/2; SB blocked p38 MAPK, but further activated Erk1/2. RT-PCR confirmed that NO center dot and SB could additively stabilize certain mRNA transcripts, an effect abolished by Erk1/2 inhibition. In affected genes, these responses were associated with CU-rich elements (CURE) in 3'-untransiated regions (3'-UTR). NO center dot stabilized the mRNA of a CURE-containing reporter gene, while repressing translation. Dominant-negative Mek1, an Erk1/2 inhibitor, abolished this effect. NO center dot similarly stabilized, but blocked translation of MAP3K7IP2, a natural CURE-containing gene. NO center dot increased hnRNP translocation to the cytoplasm and binding to CURE. Over-expression of hnRNP K, like NO center dot, repressed translation of CURE-containing mRNA. These findings define a sequence-specific mechanism of NO center dot-triggered gene regulation that stabilizes mRNA, but represses translation.