DNA Sequences in gal Operon Override Transcription Elongation Blocks
| Autor: | Mikhail Kashlev, Phuoc Le, Sankar Adhya, Natalia Komissarova, Dale E.A. Lewis |
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| Rok vydání: | 2008 |
| Předmět: |
DNA
Bacterial Transcription Genetic 5' Flanking Region Molecular Sequence Data Repressor Biology Article chemistry.chemical_compound Structural Biology Transcription (biology) RNA polymerase Operon Escherichia coli gal operon 3' Flanking Region Promoter Regions Genetic Molecular Biology Base Sequence Oligonucleotide Escherichia coli Proteins Galactose RNA Promoter DNA-Directed RNA Polymerases Gene Expression Regulation Bacterial Oligonucleotides Antisense Molecular biology RNA Bacterial chemistry DNA Transcription Factors |
| Zdroj: | Journal of Molecular Biology. 382:843-858 |
| ISSN: | 0022-2836 |
| Popis: | The DNA loop that represses transcription from galactose (gal) promoters is infrequently formed in stationary-phase cells because the concentration of the loop architectural protein HU is significantly low at that state, resulting in expression of the operon in the absence of the gal inducer D-galactose. Unexpectedly, transcription from the gal promoters under these conditions overrides physical block because of the presence of the Gal repressor bound to an internal operator (O(I)) located downstream of the promoters. We have shown here that although a stretch of pyrimidine residues (UUCU) in the RNA:DNA hybrid located immediately upstream of O(I) weakens the RNA:DNA hybrid and favors RNA polymerase (RNAP) pausing and backtracking, a stretch of purines (GAGAG) in the RNA present immediately upstream of the pause sequence in the hybrid acts as an antipause element by stabilizing the RNA:DNA duplex and preventing backtracking. This facilitates forward translocation of RNAP, including overriding of the DNA-bound Gal repressor barrier at O(I). When the GAGAG sequence is separated from the pyrimidine sequence by a 5-bp DNA insertion, RNAP backtracking is favored from a weak hybrid to a more stable hybrid. RNAP backtracking is sensitive to Gre factors, D-galactose, and antisense oligonucleotides. The ability of a native DNA sequence to override transcription elongation blocks in the gal operon uncovers a previously unknown way of regulating gal metabolism in Escherichia coli. It also explains the synthesis of gal enzymes in the absence of inducer for biosynthetic reactions. |
| Databáze: | OpenAIRE |
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