Two-Hybrid System for Characterization of Protein-Protein Interactions in
            <i>E. coli</i>

Hays, Lori B.; Chen, Yuen-Shing A.; Hu, James C.

doi:10.2144/00292st04

Cited by 22 publications

(11 citation statements)

References 27 publications

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“…sensitivity) in repression is the largest when K R1 and K R2 are equal. Similar schemes have been generalized for co-repression by two species of repressors [35][36][37], and can be used to mimic the logical NAND function [17 • ].…”

Section: Cooperative Repressionmentioning

confidence: 99%

Transcriptional regulation by the numbers: applications

Bintu

Buchler

García

et al. 2005

Current Opinion in Genetics & Development

347

457

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With the increasing amount of experimental data on gene expression and regulation, there is a growing need for quantitative models to describe the data and relate them to their respective context. Thermodynamic models provide a useful framework for the quantitative analysis of bacterial transcription regulation. This framework can facilitate the quantification of vastly different forms of gene expression from several well-characterized bacterial promoters that are regulated by one or two species of transcription factors; it is useful because it requires only a few parameters. As such, it provides a compact description useful for higher-level studies (e.g. of genetic networks) without the need to invoke the biochemical details of every component. Moreover, it can be used to generate hypotheses on the likely mechanisms of transcriptional control.

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Section: Cooperative Repressionmentioning

confidence: 99%

Transcriptional regulation by the numbers: applications

Bintu

Buchler

García

et al. 2005

Current Opinion in Genetics & Development

347

457

View full text Add to dashboard Cite

show abstract

“…Each construction was sequenced to verify its integrity. To assess the ability of cloned fragments to oligomerize, two assays were carried out, a phage sensitivity assay and ␤-galactosidase repression assay (25,26). Plasmids expressing the chimeric proteins were transformed into Escherichia coli strain AG1688, and the strains were infected with cI Ϫ .…”

Section: In Vivo Oligomerization Assaymentioning

confidence: 99%

Molecular Dissection of VirB, a Key Regulator of the Virulence Cascade of Shigella flexneri

Beloin

McKenna

Dorman

2002

Journal of Biological Chemistry

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The VirB protein is a key regulator of virulence gene expression in the facultative enteroinvasive pathogen Shigella flexneri. While genetic evidence has shown that it is required for activation of transcription of virulence genes located on a 230-kb plasmid in this bacterium, hitherto, evidence that VirB is a DNA-binding protein has been lacking. Although VirB shows extensive homology to proteins involved in plasmid partitioning, it does not resemble any known conventional transcription factor. Here we show for the first time that VirB binds to the promoter regions of the virulence genes in vivo. We also show that VirB forms dimeric and higher oligomeric structures both in vivo and in vitro and that this property is independent of DNA binding. The oligomerization activity of VirB is distributed over two domains: a leucine zipper-like motif and a carboxyl-terminal domain likely to form triple coiled structures. VirB possesses a helix-turn-helix motif, which is required for DNA binding. The amino-terminal domain of the protein is also required for DNA binding and virulence gene activation. The possibility that VirB requires a co-factor for specific interaction with target promoters in vivo is discussed.

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“…tein-protein interactions 17 . Large-scale approaches for screening protein library arrays followed by recovery and identification of the encoding gene on a massive scale will also prove useful (e.g., see ref.…”

Section: Interpromentioning

confidence: 99%

Status of genome projects for nonpathogenic bacteria and archaea

Nelson¹,

Paulsen²,

Heidelberg³

et al. 2000

Nat Biotechnol

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Since the first microbial genome was sequenced in 1995, 30 others have been completed and an additional 99 are known to be in progress. Although the early emphasis of microbial genomics was on human pathogens for obvious reasons, a significant number of sequencing projects have focused on nonpathogenic organisms, beginning with the release of the complete genome sequence of the archaeon Methanococcus jannaschii in 1996. The past 18 months have seen the completion of the genomes of several unusual organisms, including Thermotoga maritima, whose genome reveals extensive potential lateral transfer with archaea; Deinococcus radiodurans, the most radiation-resistant microorganism known; and Aeropyrum pernix, the first Crenarchaeota to be completely sequenced. Although the functional characterization of genomic data is still in its initial stages, it is likely that microbial genomics will have a significant impact on environmental, food, and industrial biotechnology as well as on genomic medicine.

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Two-Hybrid System for Characterization of Protein-Protein Interactions in E. coli

Cited by 22 publications

References 27 publications

Transcriptional regulation by the numbers: applications

Transcriptional regulation by the numbers: applications

Molecular Dissection of VirB, a Key Regulator of the Virulence Cascade of Shigella flexneri

Status of genome projects for nonpathogenic bacteria and archaea

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