The transcriptional program regulated by the tumor suppressor p53 was analyzed using oligonucleotide microarrays. A human lung cancer cell line that expresses the temperature sensitive murine p53 was utilized to quantirate mRNA levels of various genes at different time points after shifting the temperature to 32C. Inhibition of protein synthesis by cycloheximide (CHX) was used to distinguish between primary and secondary target genes regulated by p53. In the absence of CHX, 259 and 125 genes were up or down-regulated respectively; only 38 and 24 of these genes were up and down-regulated by p53 also in the presence of CHX and are considered primary targets in this cell line. Cluster analysis of these data using the super paramagnetic clustering (SPC) algorithm demonstrate that the primary genes can be distinguished as a single cluster among a large pool of p53 regulated genes. This procedure identified additional genes that co-cluster with the primary targets and can also be classified as such genes. In addition to cell cycle (e.g., p21, TGF-[3, Cyclin E) and apoptosis (e.g., Fas, Bak, IAP) related genes, the primary targets of p53 include genes involved in many aspects of cell function, including cell adhesion (e.g., Thymosin, Smoothelin), signaling (e.g., H-ras, Diacylglyerol kinase), transcription (e.g., ATF3, LISCH7), neuronal growth (e.g., Ninjurin, NSCL2) and DNA repair (e.g., BTG2, DDB2). The results suggest that p53 activates concerted opposing signals and exerts its effect through a diverse network of transcriptional changes that collectively alter the cell phenotype in response to stress.