Binding of Escherichia coli RNA polymerase to a promoter carrying mutations that stop transcription initiation.

The gal P2 promoter can be inactivated by point mutations located in the -10 hexamer sequence or immediately upstream from it. Mutations at either site reduce expression in vivo and prevent the formation, in vitro, of tight complexes with RNA polymerase that give a strong footprint and can initiate transcription. However, with a mutation upstream from the -10 region, RNA polymerase could still make a specific contact with gal promoter DNA as judged by interference with cleavage by restriction enzyme SfaNI at a site within the promoter. In contrast, with a mutation in the -10 hexamer sequence, RNA polymerase could not make this contact and does not interfere with restriction by SfaNI.