CrtJ Bound to Distant Binding Sites Interacts Cooperatively to Aerobically Repress Photopigment Biosynthesis and Light Harvesting II Gene Expression in Rhodobacter capsulatus

Abstract

Expression of light harvesting II genes and of bacteriochlorophyll and carotenoid biosynthesis genes in Rhodobacter capsulatus is repressed under aerobic growth conditions by the transcription factor CrtJ. In this study, we demonstrate that the crtA-crtI intergenic region contains divergent promoters that initiate transcription 116 base pairs apart, based on primer extension analyses. DNase I protection assays demonstrate that purified CrtJ binds to one palindrome that overlaps the crtA -10 promoter recognition sequence as well as to a second palindrome that overlaps the -35 crtI promoter recognition sequence. Similar analyses also show that the puc promoter region contains two distant CrtJ palindromes, with one near the -35 promoter recognition sequence and the other located 240 base pairs upstream. Gel mobility shift and filter retention assays indicate that CrtJ binds in a cooperative manner to these distantly separated palindromes. In vivo expression assays with puc and crtI promoter reporter plasmids further demonstrate that aerobic repression of puc and crtI expression requires both CrtJ palindromes. These in vitro and in vivo results indicate that aerobic repression of puc, crtA, and crtI expression involves cooperative interactions between CrtJ bound to distant palindromes. A DNA looping model is discussed.