The role of the transcription factor Sp1 in cell growth and survival was investigated by induced expression of its DNA-binding C-terminal fragment. Transfection of a constitutively expressed Sp1-170C plasmid construct into HeLa cells failed to produce viable clones, suggesting that this peptide interferes with cell growth. However, transfection with the muristerone A-inducible vector system produced four clones with low levels of expression of Sp1-170C. Muristerone A transiently induced higher levels of Sp1-170C, and this was accompanied by a reduced growth rate and prolongation of the S phase of the cell cycle. This is the first report that a dominant negative Sp1 can affect the cell cycle.