In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH
Abstract
Transcription of the phage-shock protein (psp) operon in Escherichia coli is driven by a sigma(54) promoter, stimulated by integration host factor and dependent on an upstream, cis-acting sequence and an activator protein, PspF. PspF belongs to the enhancer binding protein family but lacks an N-terminal regulatory domain. Purified PspF is not modified and has an ATPase activity that is increased twofold in the presence of DNA carrying the psp cis-acting sequence. Purified mutant I-Iis-tagged PspF that lacks the C-terminal DNA-binding motif has a DNA-independent ATPase activity when present at 30-fold the concentration of the wild-type protein. Both proteins oligomerize in solution in an ATP and DNA-independent manner. The wild-type activator protein, but not the DNA-binding mutant, binds specifically to the cis-acting sequence. Analysis of the sequence protected by PspF demonstrates the presence of two upstream binding sites within the sequence, UAS I and UAS II, which together constit...ute the psp enhancer. Protection at low protein concentrations is more pronounced and more extensive on a supercoiled DNA than on a linear template. Full expression of the psp operon upon hyperosmotic shock depends on wild-type PspF, but only partially requires the presence of the psp enhancer.
Keywords:
sigma(54) / PspF / PspF Delta HTH / psp enhancer / oligomerizationSource:
Journal of Molecular Biology, 1999, 285, 2, 469-483Publisher:
- Academic Press Ltd- Elsevier Science Ltd, London
DOI: 10.1006/jmbi.1998.2263
ISSN: 0022-2836
PubMed: 9878422
WoS: 000078138300005
Scopus: 2-s2.0-0033555834
Collections
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Jovanović, Goran AU - Rakonjac, J AU - Model, P PY - 1999 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/119 AB - Transcription of the phage-shock protein (psp) operon in Escherichia coli is driven by a sigma(54) promoter, stimulated by integration host factor and dependent on an upstream, cis-acting sequence and an activator protein, PspF. PspF belongs to the enhancer binding protein family but lacks an N-terminal regulatory domain. Purified PspF is not modified and has an ATPase activity that is increased twofold in the presence of DNA carrying the psp cis-acting sequence. Purified mutant I-Iis-tagged PspF that lacks the C-terminal DNA-binding motif has a DNA-independent ATPase activity when present at 30-fold the concentration of the wild-type protein. Both proteins oligomerize in solution in an ATP and DNA-independent manner. The wild-type activator protein, but not the DNA-binding mutant, binds specifically to the cis-acting sequence. Analysis of the sequence protected by PspF demonstrates the presence of two upstream binding sites within the sequence, UAS I and UAS II, which together constitute the psp enhancer. Protection at low protein concentrations is more pronounced and more extensive on a supercoiled DNA than on a linear template. Full expression of the psp operon upon hyperosmotic shock depends on wild-type PspF, but only partially requires the presence of the psp enhancer. PB - Academic Press Ltd- Elsevier Science Ltd, London T2 - Journal of Molecular Biology T1 - In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH EP - 483 IS - 2 SP - 469 VL - 285 DO - 10.1006/jmbi.1998.2263 ER -
@article{ author = "Jovanović, Goran and Rakonjac, J and Model, P", year = "1999", abstract = "Transcription of the phage-shock protein (psp) operon in Escherichia coli is driven by a sigma(54) promoter, stimulated by integration host factor and dependent on an upstream, cis-acting sequence and an activator protein, PspF. PspF belongs to the enhancer binding protein family but lacks an N-terminal regulatory domain. Purified PspF is not modified and has an ATPase activity that is increased twofold in the presence of DNA carrying the psp cis-acting sequence. Purified mutant I-Iis-tagged PspF that lacks the C-terminal DNA-binding motif has a DNA-independent ATPase activity when present at 30-fold the concentration of the wild-type protein. Both proteins oligomerize in solution in an ATP and DNA-independent manner. The wild-type activator protein, but not the DNA-binding mutant, binds specifically to the cis-acting sequence. Analysis of the sequence protected by PspF demonstrates the presence of two upstream binding sites within the sequence, UAS I and UAS II, which together constitute the psp enhancer. Protection at low protein concentrations is more pronounced and more extensive on a supercoiled DNA than on a linear template. Full expression of the psp operon upon hyperosmotic shock depends on wild-type PspF, but only partially requires the presence of the psp enhancer.", publisher = "Academic Press Ltd- Elsevier Science Ltd, London", journal = "Journal of Molecular Biology", title = "In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH", pages = "483-469", number = "2", volume = "285", doi = "10.1006/jmbi.1998.2263" }
Jovanović, G., Rakonjac, J.,& Model, P.. (1999). In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH. in Journal of Molecular Biology Academic Press Ltd- Elsevier Science Ltd, London., 285(2), 469-483. https://doi.org/10.1006/jmbi.1998.2263
Jovanović G, Rakonjac J, Model P. In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH. in Journal of Molecular Biology. 1999;285(2):469-483. doi:10.1006/jmbi.1998.2263 .
Jovanović, Goran, Rakonjac, J, Model, P, "In vivo and in vitro activities of the Escherichia coli sigma(54) transcription activator, PspF, and its DNA-binding mutant, PspF Delta HTH" in Journal of Molecular Biology, 285, no. 2 (1999):469-483, https://doi.org/10.1006/jmbi.1998.2263 . .