In vivo increase of solubility of overexpressed Hha protein by tandem expression with interacting protein H-NS

Pons, José I.; Rodriguez, Sonia; Madrid, Cristina; Juárez, Antonio; Nieto, J. M.

doi:10.1016/j.pep.2004.02.018

Cited by 7 publications

(3 citation statements)

References 40 publications

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“…Complex-To generate large quantities of Hha and H-NS for crystallographic studies, we employed a co-expression system previously described to increase the solubility properties of Hha (42). Hha was overexpressed with an N-terminal His 6 tag (Hha His6 ) from a pET15b vector in tandem with an untagged version of H-NS from pCDF1b.…”

Section: Purification and Crystallization Of The Hha/h-ns(1-46)mentioning

confidence: 99%

Structural Insights into the Regulation of Foreign Genes in Salmonella by the Hha/H-NS Complex

Ali

Whitney

Stevenson

et al. 2013

Journal of Biological Chemistry

108

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Section: Purification and Crystallization Of The Hha/h-ns(1-46)mentioning

confidence: 99%

Structural Insights into the Regulation of Foreign Genes in Salmonella by the Hha/H-NS Complex

Ali

Whitney

Stevenson

et al. 2013

Journal of Biological Chemistry

108

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“…Difficulties in obtaining a complete high-resolution structure of H-NS have thus far thwarted attempts to determine the specific mechanism of DNA binding (Dorman 2004). As Hha stabilizes the H-NS protein in vitro, perhaps someday it will be possible to obtain a complete structure of an Hha/H-NS/promoter complex (Pons et al 2004).…”

Section: Future Directionsmentioning

confidence: 99%

Silencing of xenogeneic DNA by H-NS—facilitation of lateral gene transfer in bacteria by a defense system that recognizes foreign DNA

Navarre¹,

McClelland²,

Libby³

et al. 2007

Genes Dev.

270

261

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Lateral gene transfer has played a prominent role in bacterial evolution, but the mechanisms allowing bacteria to tolerate the acquisition of foreign DNA have been incompletely defined. Recent studies show that H-NS, an abundant nucleoid-associated protein in enteric bacteria and related species, can recognize and selectively silence the expression of foreign DNA with higher adenine and thymine content relative to the resident genome, a property that has made this molecule an almost universal regulator of virulence determinants in enteric bacteria. These and other recent findings challenge the ideas that curvature is the primary determinant recognized by H-NS and that activation of H-NS-silenced genes in response to environmental conditions occurs through a change in the structure of H-NS itself. Derepression of H-NS-silenced genes can occur at specific promoters by several mechanisms including competition with sequence-specific DNA-binding proteins, thereby enabling the regulated expression of foreign genes. The possibility that microorganisms maintain and exploit their characteristic genomic GC ratios for the purpose of self/non-self-discrimination is discussed.

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“…The MBP-Hha protein was used to obtain adequate expression and to maintain solubility of Hha. 44 Figure 3(b) shows the gel mobility shift of three fragments from the hilA promoter, spanning K242 to K55, K55 to C90, and C100 to C300. A single shifted band of highly retarded H-NS-DNA complex was detected with fragments K242/K55 and C100/C300 at low concentrations of H-NS (60-240 nM).…”

Section: Action Sites Of H-ns and Hha On Hila Flanking Sequencesmentioning

confidence: 99%

Crucial Roles of Both Flanking Sequences in Silencing of the hilA promoter in Salmonella enterica

Olekhnovich

Kadner

2006

Journal of Molecular Biology

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In vivo increase of solubility of overexpressed Hha protein by tandem expression with interacting protein H-NS

Cited by 7 publications

References 40 publications

Structural Insights into the Regulation of Foreign Genes in Salmonella by the Hha/H-NS Complex

Structural Insights into the Regulation of Foreign Genes in Salmonella by the Hha/H-NS Complex

Silencing of xenogeneic DNA by H-NS—facilitation of lateral gene transfer in bacteria by a defense system that recognizes foreign DNA

Crucial Roles of Both Flanking Sequences in Silencing of the hilA promoter in Salmonella enterica

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