Stimulation of a Chlamydomonas chloroplast promoter by novobiocin in situ and in E. coli implies regulation by torsional stress in the chloroplast DNA

Abstract

We have characterized regulation of a complex Chlamydomonas reinhardtii chloroplast (PA) whose activity is stimulated by the DNA gyrase inhibitor novobiocin, both in the alga itself and in a heterologous Escherichia coli plasmid system. Since novobiocin is known to reduce torsional stress in E. coli DNA, we interpret our results to mean that PA is regulated by torsional stress in the chloroplast DNA. In E. coli, where we could readily manipulate PA, we found that this regulation depends on sequences upstream of PA. These sequences contain at least two different kinds of silencing elements that inhibit PA in the absence of novobiocin. Novobiocin stimulates PA only when the promoter-distal silencing element is present.