Abstract

Nitric oxide (NO
) can stabilize mRNA by activating p38 mitogen-activated protein kinase (MAPK). Here, transcript stabilization by NO
was investigated in human THP-1 cells using microarrays. After LPS pre-stimulation, cells were treated with actinomycin D and then exposed to NO
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
activated p38 MAPK and Erk1/2; SB blocked p38 MAPK, but further activated Erk1/2. RTPCR confirmed that NO
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 30-untranslated regions (30-UTR). NO
stabilized the mRNA of a CURE-containing reporter gene, while repressing translation. Dominant-negative Mek1, an Erk1/2 inhibitor, abolished this effect. NO
similarly stabilized, but blocked translation of MAP3K7IP2, a natural CURE-containing gene. NO
increased hnRNP translocation to the cytoplasm and binding to CURE. Over-expression of hnRNP K, like NO
, repressed translation of CURE-containing mRNA. These findings define a sequence-specific mechanism of NO
-triggered gene regulation that stabilizes mRNA, but represses translation.



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