Signal transduction via PI 3-kinases plays an important role in regulating the cellular processes of cell growth, survival, proliferation, and motility. The stimulated generation of reactive oxygen species is a necessary component of the signal transduction mechanisms by which many growth factors and cytokines activate this signaling pathway and elicit their cellular responses. Evidence now supports the oxidative inactivation of both tyrosine phosphatases acting upstream of PI 3-kinase, and of the lipid phosphatase PTEN as components of the normal stimulated regulation of PI 3-kinase signaling. However, the effects of chronic oxidative stress appear rather different, particularly a proposed role for nitrosylation of Akt and other targets leading to inhibition of PI 3-kinase signaling during diabetic insulin resistance in muscle. Recently, evidence has also begun to emerge, indicating that physiological redox signaling may display the same tight spatial and temporal specificity as seen with many other signal transduction systems in terms of targeting individual proteins for modification, and of enzymatic reversal mechanisms. This review will focus upon the details of these and other roles for reactive oxygen and nitrogen species in the regulation of PI 3-kinase signaling, both during acute stimulation and chronic oxidative stress, and the evidence for their significance.