Abstract:
:Fundamental questions in bacterial gene regulation concern how multiple regulatory proteins interact with the transcription apparatus at a single promoter and what are the roles of protein contacts with RNA polymerase and changes in DNA conformation. Transcription of the Escherichia coli uhpT gene, encoding the inducible sugar phosphate transporter, is dependent on the response regulator UhpA and is stimulated by the cyclic AMP receptor protein (CAP). UhpA binds to multiple sites in the uhpT promoter between positions -80 and -32 upstream of the transcription start site, and CAP binds to a single site centered at position -103.5. The role in uhpT transcription of portions of RNA polymerase Esigma(70) holoenzyme which affect regulation at other promoters was examined by using series of alanine substitutions throughout the C-terminal domains of RpoA (residues 255 to 329) and of RpoD (residues 570 to 613). Alanine substitutions that affected in vivo expression of a uhpT-lacZ transcriptional fusion were tested for their effect on in vitro transcription activity by using reconstituted holoenzymes. Consistent with the binding of UhpA near the -35 region, residues K593 and K599 in the C-terminal region of RpoD were necessary for efficient uhpT expression in response to UhpA alone. Their requirement was overcome when CAP was also present. In addition, residues R265, G296, and S299 in the DNA-binding surface of the C-terminal domain of RpoA (alphaCTD) were important for uhpT transcription even in the presence of CAP. Substitutions at several other positions had effects in cells but not during in vitro transcription with saturating levels of the transcription factors. Two DNase-hypersensitive sites near the upstream end of the UhpA-binding region were seen in the presence of all three transcription factors. Their appearance required functional alphaCTD but not the presence of upstream DNA. These results suggest that both transcription activators depend on or interact with different subunits of RNA polymerase, although their role in formation of proper DNA geometry may also be crucial.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Olekhnovich IN,Kadner RJdoi
10.1128/JB.181.23.7266-7273.1999keywords:
subject
Has Abstractpub_date
1999-12-01 00:00:00pages
7266-73issue
23eissn
0021-9193issn
1098-5530journal_volume
181pub_type
杂志文章abstract::The N-terminal amino acid sequence of the polyhydroxyalkanoic acid (PHA) granule-associated M(r)-15,500 protein of Rhodococcus ruber (the GA14 protein) was analyzed. The sequence revealed that the corresponding structural gene is represented by open reading frame 3, encoding a protein with a calculated M(r) of 14,175 ...
journal_title:Journal of bacteriology
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doi:10.1128/jb.176.14.4328-4337.1994
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journal_title:Journal of bacteriology
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更新日期:1973-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:2000-11-01 00:00:00
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更新日期:1969-11-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1993-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1999-02-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:2018-11-26 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.173.2.521-529.1991
更新日期:1991-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.134.1.237-245.1978
更新日期:1978-04-01 00:00:00
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更新日期:2007-01-01 00:00:00
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更新日期:1971-01-01 00:00:00
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更新日期:2005-07-01 00:00:00
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更新日期:1968-04-01 00:00:00
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更新日期:1999-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1975-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1971-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1995-06-01 00:00:00