Domain organization of RNA polymerase α subunit: C-terminal 85 amino acids constitute a domain capable of dimerization and DNA binding

Blatter, Erich E.; Ross, Wilma; Tang, Hong; Gourse, Richard L.; Ebright, Richard H.

doi:10.1016/s0092-8674(94)90682-3

Cited by 232 publications

(226 citation statements)

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“…The fact that a 230þ3 and a 235þ4 are inefficiently associated and form predominantly homodimers (cf. the efficient association of a 255 and heterodimer formation) supports the role of the C-terminal segment in subunit dimerization (Blatter et al 1994). Furthermore, coprecipitation also confirms that the extreme C-terminus of a in holoenzyme is exposed ( Fig.…”

Section: Stability and Associationsupporting

confidence: 62%

“…It is interesting to note that the truncated a derivatives were not processed any closer to the N-terminus. The interdomain linker of the isolated subunit (c. residues 235¹250) has been shown to be susceptible to proteolysis in vitro (Blatter et al 1994;Negishi et al 1995). It could be argued that the absence of any cleavage in this region in vivo by E. coli protease(s) suggests that the interdomain linker is protected from proteolysis upon assembly into core/ holoenzyme.…”

Section: Stability and Associationmentioning

confidence: 99%

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A nested set of C‐terminal deletions of the α subunit of Escherichia coli RNA polymerase define regions concerned with assembly, proteolysis, stabilization and transcriptional activation in vivo

et al. 1997

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Section: Stability and Associationsupporting

confidence: 62%

Section: Stability and Associationmentioning

confidence: 99%

A nested set of C‐terminal deletions of the α subunit of Escherichia coli RNA polymerase define regions concerned with assembly, proteolysis, stabilization and transcriptional activation in vivo

et al. 1997

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“…The ␣ subunit contains an N-terminal domain (␣NTD; residues 8 to 232) and a C-terminal domain (␣CTD; residues 249 to 329) (5, 21, 44). ␣NTD and ␣CTD are separated by a flexible linker that is at least 13 amino acids long (5,20,25). ␣CTD plays important roles in transcription activation by contacting a number of transcription factors (6, 18) or through interacting with an AϩT-rich DNA sequence referred to as the UP element (16).…”

mentioning

confidence: 99%

The UP Element Is Necessary but Not Sufficient for Growth Rate-Dependent Control of the Escherichia coli guaB Promoter

Husnain

Thomas

2008

J Bacteriol

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The Escherichia coli guaB promoter (P guaB ) regulates the transcription of two genes, guaB and guaA, that are required for de novo synthesis of GMP, a precursor for the synthesis of guanine nucleoside triphosphates. The activity of P guaB is subject to growth rate-dependent control (GRDC). Here we show that the A؉T-rich sequence located between positions ؊59 and ؊38 relative to the guaB transcription start site stimulates transcription from P guaB ϳ8-to 10-fold and, in common with other UP elements, requires the C-terminal domain of the RNA polymerase ␣ subunit for activity. Like the rrnB P1 UP element, the P guaB UP element contains two independently acting subsites located at positions ؊59 to ؊47 and ؊46 to ؊38 and can stimulate transcription when placed upstream of the lacP1 promoter. We reveal a novel role for the P guaB UP element by demonstrating that it is required for GRDC. The involvement of the UP element in GRDC also requires the participation of sequences located at least 100 bp upstream of the guaB transcription start site. These sequences are required for down-regulation of P guaB activity at lower growth rates.Escherichia coli RNA polymerase (RNAP) is a multisubunit complex consisting of an ␣ subunit homodimer, single ␤, ␤Ј, and subunits, and one of several subunits (14). The ␣ subunit contains an N-terminal domain (␣NTD; residues 8 to 232) and a C-terminal domain (␣CTD; residues 249 to 329) (5, 21, 44). ␣NTD and ␣CTD are separated by a flexible linker that is at least 13 amino acids long (5,20,25). ␣CTD plays important roles in transcription activation by contacting a number of transcription factors (6, 18) or through interacting with an AϩT-rich DNA sequence referred to as the UP element (16).UP elements stimulate transcription by recruiting ␣CTD to DNA (32, 43). The best-studied UP element spans the region from Ϫ59 to Ϫ38 at the rrnB P1 promoter (16). The rrnB P1 UP element contains two domains, a ϳ9-bp proximal UP element subsite located at positions Ϫ46 to Ϫ38 and a ϳ13-bp distal UP element subsite located at positions Ϫ59 to Ϫ47. Each subsite functions independently by recruiting one ␣CTD (11). The amino acid residues in ␣CTD that comprise the 265 determinant, including residues V264, R265, N268, N294, G296, K298, and S299, interact with UP element and A-tract sequences by contacting the DNA backbone, and R265 makes additional contacts with bases in the minor groove (4, 32, 43). These residues are the most important for rrnB P1 UP element utilization both in vivo and in vitro (13, 35). In the absence of a strong distal UP element subsite, ␣CTD bound to a consensus or near-consensus proximal UP element subsite also makes productive contacts with region 4.2 of 70 that serve to stimulate transcription (7,35).The E. coli guaB promoter (P guaB ) regulates the transcription of two genes, guaB and guaA, which together constitute the guaBA operon. The guaB and guaA genes encode IMP dehydrogenase and GMP synthetase, respectively, and are required for the biosynthesis of GMP from IMP (22, 42). P guaB...

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“…Results of deletion analysis (Kimura et al 1994) and limited proteolysis studies (Blatter et al 1994;Negishi et al 1995) suggested that this region forms a compact globular domain. In order to identify the subunitsubunit contact sites, we generated a set of 11 -insertion mutants containing an alanine-serine (AS) dipeptide at 20 amino acid intervals within this domain and analysed their activities of enzyme assembly in vitro (Kimura & Ishihama 1995a).…”

Section: Introductionmentioning

confidence: 99%

**Subunit assembly in vivo of Escherichia coli RNA polymerase: role of the amino‐terminal assembly domain of alpha subunit**

Kimura

Ishihama

1996

Genes to Cells

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Domain organization of RNA polymerase α subunit: C-terminal 85 amino acids constitute a domain capable of dimerization and DNA binding

Cited by 232 publications

References 31 publications

A nested set of C‐terminal deletions of the α subunit of Escherichia coli RNA polymerase define regions concerned with assembly, proteolysis, stabilization and transcriptional activation in vivo

A nested set of C‐terminal deletions of the α subunit of Escherichia coli RNA polymerase define regions concerned with assembly, proteolysis, stabilization and transcriptional activation in vivo

The UP Element Is Necessary but Not Sufficient for Growth Rate-Dependent Control of the Escherichia coli guaB Promoter

**Subunit assembly in vivo of Escherichia coli RNA polymerase: role of the amino‐terminal assembly domain of alpha subunit**

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