The effects of covalent additions of a psoralen on transcription by E. coli RNA polymerase.

Synthetic DNA substrates containing a site-specifically engineered psoralen monoadduct or diadduct were used to characterize the response of the E. coli RNA polymerase elongation complex to these lesions. The psoralen derivative HMT (4'-hydroxymethyl-4,5', 8-trimethylpsoralen) was site specifically placed into two synthetic double-stranded DNA fragments each of which contained an E. coli RNA polymerase promoter at one end. The HMT molecule was attached to the middle of the DNA fragments as either a furan-side monoadduct or an interstrand diadduct. Transcription off the HMT crosslinked DNA templates showed that E. coli RNA polymerase terminated at the HMT diadduct site, i. e., one nucleotide before the modified thymidine residue on the template strand. The furan-side monoadduct when on the template strand also blocked transcription by the polymerase. However, no effect on transcription was observed when the monoadduct was located on the non-template strand.