The protooncogene p21, a monomeric G protein family member, plays a critical role in converting extracellular signals into intracellular biochemical events. Here, we report that nitric oxide (NO) activates p21 in human T cells as evidenced by an increase in GTP-bound p21. In vitro studies using pure recombinant p21 demonstrate that the activation is direct and reversible. Circular dichroism analysis reveals that NO induces a profound conformational change in p21 in association with GDP/GTP exchange. The mechanism of activation is due to S-nitrosylation of a critical cysteine residue which stimulates guanine nucleotide exchange. Furthermore, we demonstrate that p21 is essential for NO-induced downstream signaling, such as NF-B activation, and that endogenous NO can activate p21 in the same cell. These studies identify p21 as a target of NO in T cells and suggest that NO activates p21 by an action which mimics that of guanine nucleotide exchange factors.

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