Heat shock protein 60 and heat shock protein 10 are mitochondrial chaperonin proteins. Here, we report that the expressions of heat shock protein 60 and heat shock protein 10 were upregulated in interferon-gamma-treated C6 astroglioma cells, and the 582 bp in the bidirectional promoter of the heat shock protein 60 and heat shock protein 10 genes is responsible for interferon-gamma-induced induction. The induction of heat shock protein 60 and heat shock protein 10 by interferon-gamma was virtually abolished by introducing mutations into the putative signal transducers and activators of transcription 3-response element in the promoter, and the same mutation impaired increment of the signal transducers and activators of transcription 3-binding after interferon-gamma treatment. Moreover, Rac1 GTPase was required for maximal heat shock protein 10 and heat shock protein 60 inductions by interferon-gamma. These results suggest that interferon-gamma-induced upregulations of heat shock protein 60 and heat shock protein 10 in C6 astroglioma cells are mediated by the signal transducers and activators of transcription 3-binding site, localized in the bidirectional promoter.