High resolution mapping of E.coli transcription elongation complex in situ reveals protein interactions with the non-transcribed strand.
We have used chemical probes and UV light to perform a high resolution mapping of an Escherichia coli transcription elongation complex that was arrested in vivo by a protein readblock at a position distal to the promoter. The in situ probing data provide a precise picture of a constrained ternary complex in which the front edge of the polymerase is located at <6 bp from the catalytic center. Furthermore, our analyses reveal protein contacts with the non-transcribed strand within the arrested ternary complex. Thus, these results contribute substantially to the emerging view of a flexible transcription elongation complex in which the non-transcribed strand is an important regulatory element.
Guérin
M
M
Centre de Biophysique Moléculaire, CNRS, Orléans, France.
Leng
M
M
Rahmouni
A R
AR
D000079:Acetaldehyde / Q000031:analogs & derivatives / Q000494:pharmacology; D001426:Bacterial Proteins; D001483:Base Sequence; D018983:DNA Footprinting; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D007763:Lac Operon / Q000235:genetics; D056968:Lac Repressors; D008969:Molecular Sequence Data; D009993:Osmium Tetroxide / Q000494:pharmacology; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012097:Repressor Proteins; D013463:Sulfuric Acid Esters / Q000494:pharmacology; D013728:Terminator Regions, Genetic / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*; D018911:Trinucleotide Repeats; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012097:Repressor Proteins; D013463:Sulfuric Acid Esters; C004656:chloroacetaldehyde; D012321:DNA-Directed RNA Polymerases; D000079:Acetaldehyde; C007482:dimethyl sulfate; D009993:Osmium Tetroxide