Streptococcal Reporter Gene-Fusion Vector for Identification of in Vivo Expressed Genes

Kılıç, Ali; Herzberg, Mark C.; Meyer, Mary Hockenberry; Zhao, Xuemei; Lin, Tao

doi:10.1006/plas.1999.1408

Cited by 45 publications

(48 citation statements)

References 28 publications

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“…Later, the cat gene was also used in IVET studies of Shigella flexneri (14), Salmonella enterica serovar Typhimurium (98,157), Helicobacter pylori (8), Yersinia enterocolitica (300), Yersinia ruckeri (65), Streptococcus gordonii (127), Escherichia coli (126), and Burkholderia pseudomallei (239). Bacteria harboring promoters that are specifically induced in the wild were selected by administrating chloramphenicol to the host.…”

Section: Selection Strategies In Ivetmentioning

confidence: 99%

“…69, 2005 NICHE-SPECIFIC GENE EXPRESSION 247 Besides transcriptional gene regulation, some host-induced genes are involved in posttranslational regulation. IVET enabled the identification of serine/threonine protein kinases that are specifically expressed in Pseudomonas aeruginosa (290), Lactobacillus plantarum (24), and Streptococcus gordonii (127) during interaction with mouse and rabbit hosts. Serine/threonine kinases are ubiquitous in both prokaryotic and eukaryotic cells and are thought to play a central role in signal transduction.…”

Section: Regulatory Genesmentioning

confidence: 99%

“…For instance, the IVET library constructed in Streptococcus gordonii was originally used to study gene expression during infection of mice (127). The same library was subsequently used to analyze gene expression during growth on saliva-coated hydroxyapatite (142) and during biofilm formation on polystyrene surfaces (304).…”

Section: Ivet: a Powerful And Flexible Toolmentioning

confidence: 99%

See 2 more Smart Citations

Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression

Rediers

Rainey

Vanderleyden

et al. 2005

Microbiol Mol Biol Rev

139

120

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A major challenge for microbiologists is to elucidate the strategies deployed by microorganisms to adapt to and thrive in highly complex and dynamic environments. In vitro studies, including those monitoring genomewide changes, have proven their value, but they can, at best, mimic only a subset of the ensemble of abiotic and biotic stimuli that microorganisms experience in their natural habitats. The widely used gene-to-phenotype approach involves the identification of altered niche-related phenotypes on the basis of gene inactivation. However, many traits contributing to ecological performance that, upon inactivation, result in only subtle or difficult to score phenotypic changes are likely to be overlooked by this otherwise powerful approach. Based on the premise that many, if not most, of the corresponding genes will be induced or upregulated in the environment under study, ecologically significant genes can alternatively be traced using the promoter trap techniques differential fluorescence induction and in vivo expression technology (IVET). The potential and limitations are discussed for the different IVET selection strategies and system-specific variants thereof. Based on a compendium of genes that have emerged from these promoter-trapping studies, several functional groups have been distinguished, and their physiological relevance is illustrated with follow-up studies of selected genes. In addition to confirming results from largely complementary approaches such as signature-tagged mutagenesis, some unexpected parallels as well as distinguishing features of microbial phenotypic acclimation in diverse environmental niches have surfaced. On the other hand, by the identification of a large proportion of genes with unknown function, these promoter-trapping studies underscore how little we know about the secret lives of bacteria and other microorganisms

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Section: Selection Strategies In Ivetmentioning

confidence: 99%

Section: Regulatory Genesmentioning

confidence: 99%

Section: Ivet: a Powerful And Flexible Toolmentioning

confidence: 99%

See 1 more Smart Citation

Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression

Rediers

Rainey

Vanderleyden

et al. 2005

Microbiol Mol Biol Rev

139

120

View full text Add to dashboard Cite

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“…This group is widely distributed, and the members occur in both pathogenic and commensal species and include Rgg of Streptococcus gordonii, which is required for extracellular glucosyltransferase expression (29,30); GadR of Lactococcus lactis, which is required for glutamate-dependent acid tolerance (27); MutR, which is required for expression of the mutacin lantibiotic, MutA, of Streptococcus mutans (23); and the plasmid-encoded LasX protein of Lactobacillus sakei, which regulates the synthesis of and immunity to the lantibiotic lactocin S (25,28). Additional uncharacterized Rgg-like proteins are encoded by the genomes of Streptococcus pneumoniae (31), Streptococcus agalactiae (13), Streptococcus oralis (10), Streptococcus sanguis (34), Streptococcus equi (http://www.sanger.ac.uk), and Listeria monocytogenes (12), and some genomes, like those of S. pyogenes (9), S. gordonii (15,33), S. pneumoniae (31), and S. mutans (1), contain multiple rgg-like genes.…”

mentioning

confidence: 99%

Contribution of Invariant Residues to the Function of Rgg Family Transcription Regulators

Loughman

Caparon

2007

J Bacteriol

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The Rgg family of transcription regulators is widely distributed among gram-positive bacteria, yet how these proteins control transcription is poorly understood. Using Streptococcus pyogenes RopB as a model, we demonstrated that residues invariant among Rgg-like regulators are critical for function and obtained evidence for a mechanism involving protein complex formation.The Rgg-like regulators constitute a conserved family of proteins that modulate transcription in gram-positive bacteria. This group is widely distributed, and the members occur in both pathogenic and commensal species and include Rgg of Streptococcus gordonii, which is required for extracellular glucosyltransferase expression (29, 30); GadR of Lactococcus lactis, which is required for glutamate-dependent acid tolerance (27); MutR, which is required for expression of the mutacin lantibiotic, MutA, of Streptococcus mutans (23); and the plasmid-encoded LasX protein of Lactobacillus sakei, which regulates the synthesis of and immunity to the lantibiotic lactocin S (25, 28). Additional uncharacterized Rgg-like proteins are encoded by the genomes of Streptococcus pneumoniae (31), Streptococcus agalactiae (13), Streptococcus oralis (10), Streptococcus sanguis (34), Streptococcus equi (http://www.sanger.ac.uk), and Listeria monocytogenes (12), and some genomes, like those of S. pyogenes (9), S. gordonii (15, 33), S. pneumoniae (31), and S. mutans (1), contain multiple rgg-like genes.How the members of this extensive family function to regulate gene expression is not well understood. Rgg-like proteins have a helix-turn-helix motif in the amino terminus of the polypeptide, which is a conserved DNA-binding domain found in other families of transcription regulators (17). Only recently has it been established that any Rgg-like proteins bind specifically to DNA to regulate transcription. For example, association with nucleic acid has been demonstrated for Rgg of S. gordonii (35), RopB (21), and LasX (24), but there is only a weak consensus binding site (24). The absence of a conserved regulatory motif in the promoter regions of genes regulated by Rgg-like proteins and the functional diversity of the regulated gene products suggest that Rgg-like proteins interact with additional regulatory networks to alter gene expression. Experimental data supporting this hypothesis were obtained in an analysis of the speB regulatory program in S. pyogenes, where RopB is necessary but not sufficient for activation of transcription (21) and may influence gene expression via its ability to influence the expression of other regulators (5). The integration of Rgg pathways with other regulatory pathways could also be established through protein-protein interactions. For example, RopB has been shown to associate with a negative regulator, LacD.1, which may be a mechanism for maintaining temporally controlled expression programs (16a).Although the members of the Rgg family have been adapted to individual regulatory programs, it is likely that these proteins have a common structure ...

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“…Members of the rgg-like family have been identified in the genomes of S. pyogenes (8,27), S. pneumoniae (40), S. agalactiae (17), S. mutans (32), S. oralis (15), S. sanguis (42), S. equi (M. N. Neely and M. Caparon, unpublished data), and Listeria monocytogenes (16). Some genomes, including those of S. pyogenes (13,27) and S. gordonii (25,41), contain multiple family members, and rgg-like genes have also been found in nonpathogens as well, including Lactococcus lactis (35) and Lactobacillus sakei (33). How the members of this extensive family function to regulate gene expression is not understood.…”

mentioning

confidence: 99%

Role of RopB in Growth Phase Expression of the SpeB Cysteine Protease ofStreptococcus pyogenes

et al. 2003

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The Rgg family of transcription regulators is widely distributed among gram-positive bacteria; however, how the members of this family control transcription is poorly understood. In the pathogen Streptococcus pyogenes, the Rgg family member RopB is required for transcription of the gene that encodes the secreted SpeB cysteine protease. Expression of the protease follows distinct kinetics that involves control of transcription in response to the growth phase. In this study, the contribution of RopB to growth phase control was examined. The gene encoding the protease (speB) and ropB are transcribed divergently from a 940-bp intergenic region. Primer extension analyses, in conjunction with reporter fusion studies, revealed that the major region controlling the transcription of both speB and ropB is adjacent to ropB and that the promoters for the two genes likely overlap. Furthermore, it was found that RopB is a DNA-binding protein that specifically binds to sequences in this control region. The interrelationship between ropB and speB expression was further reflected in the observation that transcription of ropB itself is subject to growth phase control. However, while expression of ropB from a promoter expressed during the early logarithmic phase of growth could complement a ropB deletion mutant, ectopic expression of ropB did not uncouple the expression of speB from its growth phase signal. These data implicate other factors in growth phase control and suggest that regulation of ropB expression itself is not the central mechanism of control.

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Streptococcal Reporter Gene-Fusion Vector for Identification of in Vivo Expressed Genes

Cited by 45 publications

References 28 publications

Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression

Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression

Contribution of Invariant Residues to the Function of Rgg Family Transcription Regulators

Role of RopB in Growth Phase Expression of the SpeB Cysteine Protease ofStreptococcus pyogenes

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