RESUMO
Investigations into the regulation of heterotrimeric GTP-binding protein alpha-subunits in models of tumour necrosis factor-alpha (TNF)-induced cell death, revealed the selective down-regulation of the G(q)alpha/G11alpha family of G-proteins. The human HeLa and murine L929 cells treated with recombinant human TNF for up to 24 h displayed down-regulated G(q)alpha/G11alpha family protein levels, but not G(s)alpha, G(i)alpha and G(o)alpha protein levels as determined by Western analyses. This effect of TNF was observed in a concentration--and time-dependent manner, consistent with the profiles of TNF-induced cell death observed. Moreover, the functioning of G(q)alpha/G11alpha family proteins were found to be impaired in TNF-treated cells, as measured by agonist-induced [Ca2+]i release. In contrast, G(s)alpha activity was unaltered by TNF-treatment, determined by measurement of agonist-induced intracellular cyclic AMP generation. These findings in TNF-induced cytotoxic models, indicate a novel 'cross-talk' mechanism by which TNF alters Ca2+-signalling mechanisms, which may contribute towards the apoptotic and necrotic cell death.