The
protein kinase B/Akt
serine/threonine kinase, located downstream of
phosphoinositide 3-kinase (PI-3K), is a major regulator of cellular survival and proliferation.
Atypical protein kinase C (aPKC) family members are activated by
PI-3K and also contribute to cell proliferation, suggesting that Akt and aPKC might interact to activate signalling through the
PI-3K cascade. Here we demonstrate that blocking PKC activity in MDA-MB-468
breast cancer cells increased the phosphorylation and activity of Akt. Functional
PI-3K was required for the PKC inhibitors to increase Akt phosphorylation and activation, potentially owing to the activation of specific PKC
isoforms by
PI-3K. The concentration dependence of the action of the PKC inhibitors implicates aPKC in the inhibition of Akt phosphorylation and activity. In support of a role for aPKC in the regulation of Akt, Akt and PKCzeta or PKClambda/iota were readily co-precipitated from the BT-549
breast cancer cell line. Furthermore, the overexpression of PKCzeta inhibited
growth-factor-induced increases in Akt phosphorylation and activity. Thus PKCzeta associates physically with Akt and decreases Akt phosphorylation and
enzyme activity. The effects of PKC on Akt were transmitted through the
PI-3K cascade as indicated by changes in
p70 s6 kinase (p70(s6k)) phosphorylation. Thus PKCzeta, and potentially other PKC
isoenzymes, regulate
growth-factor-mediated Akt phosphorylation and activation, which is consistent with a generalized role for PKCzeta in limiting
growth factor signalling through the PI-3K/Akt pathway.