Sequence determinants of promoter activity

Abstract

The bacteriophage P22 promoter for the antirepressor (ant) gene, Pant, in the absence of Arc repressor, directs the synthesis of extremely high levels of antirepressor. Overproduction of antirepressor leads secondarily to the failure to produce progeny phage upon lytic infection. A substantial fraction of revertants of P22 arc-amber phage are pseudorevertants that have acquired additional mutations that decrease the activity of the ant promoter. DNA sequence analysis of 72 independent Pant "promoter-down" mutations reveals more than 25 different alterations that define two regions critical for promoter activity. With few exceptions, these promoter-down mutations decrease the homology of Pant with the consensus promoter sequence, demonstrating that the conserved features among a large number of different wild-type promoters are the determinants of promoter strength.l In general, different substitution mutations at the same site within the promoter have similar effects, resulting in either a severe or a mild reduction in promoter activity.