rpoZ, encoding the omega subunit of Escherichia coli RNA polymerase, is in the same operon as spoT.
Highly purified Escherichia coli RNA polymerase contains a small subunit termed omega. This subunit consists of 91 amino acids with a molecular weight of 10,105. We previously reported the cloning and sequencing of the gene encoding omega, which we call rpoZ (D. R. Gentry and R. R. Burgess, Gene 48:33-40, 1986). We constructed an rpoZ insertion mutation by placing a kanamycin resistance cassette into the coding region of the rpoZ gene. Purified RNA polymerase from strains carrying this mutation lacked detectable omega. We found that the insertion mutation conferred a slow-growth phenotype when introduced into most strains. We mapped the position of rpoZ on the E. coli chromosome by genetic techniques and by examining the restriction map of the whole chromosome and found that rpoZ maps around 82 min, very close to spoT. We determined that the slow-growth phenotype of the insertion mutant is suppressed in relA mutants and that the rpoZ insertion results in a classical SpoT- phenotype. This finding strongly suggests that rpoZ is upstream of spoT in the same operon and that the slow-growth phenotype elicited by the insertion mutation is due to polarity on spoT.
Gentry
D R
DR
McArdle Laboratory for Cancer Research, University of Wisconsin-Madison 53706.
Burgess
R R
RR
D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005838:Genotype; D046911:Macromolecular Substances; D009154:Mutation; D009876:Operon; D010641:Phenotype; D010957:Plasmids
D046911:Macromolecular Substances; D012321:DNA-Directed RNA Polymerases