Early transcribed sequences affect termination efficiency of Escherichia coli RNA polymerase.
We have constructed novel transcription templates in which we have fused the late gene promoters of Escherichia coli phages lambda and 82 upstream from three different rho-independent transcription terminators. Using an in vitro transcription assay and an in vivo galactokinase expression assay, we find that the initial portion of the transcribed region significantly affects the efficiency of some downstream terminators. We have identified, by deletion, substitution and point mutation analysis, sequences responsible for these increased levels of factor-independent readthrough. Since these important sequences occur within about 30 nucleotides of the RNA start site, we suggest that the initial portion of the transcript can affect termination efficiency.
Goliger
J A
JA
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853.
Yang
X J
XJ
Guo
H C
HC
Roberts
J W
JW
D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004926:Escherichia coli; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000235:genetics; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D012329:RNA, Bacterial; D012321:DNA-Directed RNA Polymerases