Cytoplasmic RNA Polymerase in Escherichia coli.
To obtain an estimate for the concentration of free functional RNA polymerase in the bacterial cytoplasm, the content of RNA polymerase beta and beta' subunits in DNA-free minicells from the minicell-producing Escherichia coli strain chi925 was determined. In bacteria grown in Luria-Bertani medium at 2.5 doublings/h, 1.0% of the total protein was RNA polymerase. The concentration of cytoplasmic RNA polymerase beta and beta' subunits in minicells produced by this strain corresponded to about 17% (or 2.5 microM) of the value found in whole cells. Literature data suggest that a similar portion of cytoplasmic RNA polymerase subunits is in RNA polymerase assembly intermediates and imply that free functional RNA polymerase can form a small percentage of the total functional enzyme in the cell. On infection with bacteriophage T7, 20% of the minicells produced progeny phage, whereas infection in 80% of the cells was abortive. RNA polymerase subunits in lysozyme-freeze-thaw lysates of minicells were associated with minicell envelopes and were without detectable activity in an in vitro transcription assay. Together, these results suggest that most functional RNA polymerase is associated with the DNA and that little if any segregates into DNA-free minicells.
Department of Molecular and Cell Biology, University of Texas at Dallas, Richardson, Texas 75083-0688, USA.
N
N
Shepherd
P
P
Dennis
H
H
Bremer
D017123:Bacteriophage T7 / Q000502:physiology; D003593:Cytoplasm / Q000201:enzymology*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development / Q000821:virology; D010957:Plasmids / Q000235:genetics
D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit