Effects of the antiterminator BoxA on transcription elongation kinetics and ppGpp inhibition of transcription elongation in Escherichia coli.
It has been shown previously that two different mRNA chains (lacZ and infB) are elongated at a rate of approximately 40 nucleotides (nt)/s during steady state growth on minimal medium and that the rate of mRNA chain elongation is inhibited by ppGpp in vivo. On the other hand, it was found that a truncated ribosomal RNA chain was elongated at a rate of approximately 80 nt/s, independent of growth condition (Vogel, U., and Jensen, K. F. (1994) J. Biol. Chem. 269, 16236-16241). We reasoned that the different transcriptional behavior of mRNA genes and rRNA operons might be caused by the antiterminator sequences present in the rRNA operons. To test this possibility, we have (a) inserted the minimal antiterminator boxA sequence between the promoter and the lacZ and infB genes and (b) deleted the antiterminator sequences from the rRNA transcription unit and measured transcription elongation rates in vivo on the resulting hybrid genes. We found that insertion of boxA in front of the coding region of lacZ increased the transcription elongation rate from 42 nt/s to 69 nt/s during steady state growth and that it eliminated the ppGpp-dependent decrease in the transcription elongation rate during the stringent response. On the other hand, deletion of the antiterminator sequences from the rRNA operon resulted in a reduced transcription elongation rate, but the elongation rate was still insensitive to changes in the ppGpp pool. These results are consistent with the hypothesis that the antiterminator boxA is a primary determinant of the rate of transcription elongation rate.
Vogel
U
U
Department of Biological Chemistry, University of Copenhagen, Denmark.
Jensen
K F
KF
D001483:Base Sequence; D004790:Enzyme Induction; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D006159:Guanosine Tetraphosphate / Q000494:pharmacology*; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000235:genetics; D039881:Prokaryotic Initiation Factor-2; D014176:Protein Biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D012335:RNA, Ribosomal / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D010448:Peptide Initiation Factors; D039881:Prokaryotic Initiation Factor-2; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D006159:Guanosine Tetraphosphate; D001616:beta-Galactosidase