Chlamydia trachomatis σ 28 recognizes the fliC promoter of Escherichia coli and responds to heat shock in chlamydiae

Shen, Li; Li, Maixiang; Zhang, You-Xun

doi:10.1099/mic.0.26734-0

Cited by 21 publications

(37 citation statements)

References 56 publications

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“…We also tested the possibility that RsbW was a negative regulator of C. trachomatis 28 , as had been suggested previously (39). However, we found that RsbW did not inhibit C. trachomatis 28 in S. enterica serovar Typhimurium.…”

Section: Discussionmentioning

confidence: 68%

“…Chlamydia are also difficult to propagate, and construction of stable transformants has been difficult (43). Although C. trachomatis is nonmotile, its 28 homolog was shown to recognize the fliC promoter of Escherichia coli in vitro (39). In this study, we present a genetic characterization of 28 from A. aeolicus and C. trachomatis in S. enterica serovar Typhimurium and an isolation of A. aeolicus 28 /FlgM through complementation and mutagenesis experiments.…”

Section: New England Biolabsmentioning

confidence: 99%

“…(44). Interestingly, a P araB -C. trachomatis rspD ϩ plasmid did not restore motility in E. coli with a fliA deletion (39). Although E. coli and S. enterica serovar Typhimurium possess nearly identical flagellar systems (25), the dramatic differences in complementation for motility with P araB -C. trachomatis rspD ϩ demonstrates that differences in the flagellar regulons between the two species can be uncovered.…”

Section: A Aeolicus and C Trachomatismentioning

confidence: 99%

“…These findings show inhibition of A. aeolicus 28 28 ) were constructed in motA-lac and fliC-lac backgrounds. C. trachomatis RsbW, the putative negative regulator of RspD (39), was encoded in trans by plasmid pJK627. However, there was no difference in the relative ␤-galactosidase levels in the presence of RsbW or in its absence (Fig.…”

Section: A Aeolicus and C Trachomatismentioning

confidence: 99%

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Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Karlinsey

Hughes

2006

J Bacteriol

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In Salmonella enterica serovar Typhimurium, 28 and anti-sigma factor FlgM are regulatory proteins crucial for flagellar biogenesis and motility. In this study, we used S. enterica serovar Typhimurium as an in vivo heterologous system to study 28 and anti-28 interactions in organisms where genetic manipulation poses a significant challenge due to special growth requirements. The chromosomal copy of the S. enterica serovar Typhimurium 28 structural gene fliA was exchanged with homologs of Aquifex aeolicus (an extreme thermophile) and Chlamydia trachomatis (an obligate intracellular pathogen) by targeted replacement of a tetRA element in the fliA gene location using -Red-mediated recombination. The S. enterica serovar Typhimurium hybrid strains showed 28 -dependent gene expression, suggesting that 28 activities from diverse species are preserved in the heterologous host system. A. aeolicus mutants defective for 28 /FlgM interactions were also isolated in S. enterica serovar Typhimurium. These studies highlight a general strategy for analysis of protein function in species that are otherwise genetically intractable and a straightforward method of chromosome restructuring using -Red-mediated recombination.

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Section: Discussionmentioning

confidence: 68%

Section: New England Biolabsmentioning

confidence: 99%

Section: A Aeolicus and C Trachomatismentioning

confidence: 99%

Section: A Aeolicus and C Trachomatismentioning

confidence: 99%

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Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Karlinsey

Hughes

2006

J Bacteriol

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“…This gene encodes a histonelike protein, which is believed to be involved in the conversion of the RB to EB forms of Chlamydia (5). Transcription of the rpsD gene, which encodes 28 Ct , is heat responsive (49). Thus, 28 Ct may play a role in controlling the differentiation of Chlamydia and its adaptation response during adverse environmental conditions.…”

mentioning

confidence: 99%

Selective Promoter Recognition by Chlamydial σ²⁸Holoenzyme

et al. 2006

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The transcription factor confers the promoter recognition specificity of RNA polymerase (RNAP) in eubacteria. Chlamydia trachomatis has three known sigma factors, 66 Chlamydia trachomatis is an obligate intracellular bacterial pathogen of humans. It is the major cause of bacterial sexually transmitted diseases in the United States and the leading cause of preventable blindness worldwide (47). A unique developmental cycle, characterized by the conversion of a metabolically inactive but infectious elementary body (EB) to an intracellularly replicating but noninfectious reticulate body (RB) and the return to the EB form at the end of the cycle, makes Chlamydia distinct from other intracellular bacteria (37). Little is known about how gene expression is regulated in Chlamydia, but some control is effected at the level of transcription by factors and other transcriptional regulators (1,16,39). By binding to the RNA polymerase (RNAP) core enzyme (composed of subunits ␣ 2 ␤␤Ј), the factors confer on the enzyme the ability to recognize specific DNA elements (promoters) located upstream of the transcription start site and interact with another transcriptional regulator(s) (10,15,20,57 Studies of molecular mechanisms in Chlamydia are challenging due to the difficulty of propagation and the inability to genetically manipulate this intracellular bacterium. As a result, the promoter specificities of the chlamydial factors have been investigated only in vitro, and only a few chlamydial promoters have been characterized (9,16,33,50,53,58). It is presumed that 66 , a homologue of E. coli 70

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New ex vivo reporter assay system reveals that σ factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants

2013

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Analysis of the environmental regulation of bacterial gene expression is important for understanding the nature, pathogenicity, and infection route of many pathogens. “Candidatus Phytoplasma asteris”, onion yellows strain M (OY-M), is a phytopathogenic bacterium that is able to adapt to quite different host environments, including plants and insects, with a relatively small ∼850 kb genome. The OY-M genome encodes two sigma (σ) factors, RpoD and FliA, that are homologous to Escherichia coli σ70 and σ28, respectively. Previous studies show that gene expression of OY-M dramatically changes upon the response to insect and plant hosts. However, very little is known about the relationship between the two σ factors and gene regulatory systems in OY-M, because phytoplasma cannot currently be cultured in vitro. Here, we developed an Escherichia coli-based ex vivo reporter assay (EcERA) system to evaluate the transcriptional induction of phytoplasmal genes by the OY-M-derived σ factors. EcERA revealed that highly expressed genes in insect and plant hosts were regulated by RpoD and FliA, respectively. We also demonstrated that rpoD expression was significantly higher in insect than in plant hosts and fliA expression was similar between the hosts. These data indicate that phytoplasma-derived RpoD and FliA play key roles in the transcriptional switching mechanism during host switching between insects and plants. Our study will be invaluable to understand phytoplasmal transmission, virulence expression in plants, and the effect of infection on insect fitness. In addition, the novel EcERA system could be broadly applied to reveal transcriptional regulation mechanisms in other unculturable bacteria.Phytoplasma, an unculturable plant pathogen, could infect plant and insect cells, and dramatically changes their genes upon the response to these hosts. By a new system developed in this study, interactions between phytoplasma promoters and sigma factors were analyzed, and overall gene expression regulation mechanism could be revealed. This model illustrates the RpoD and FliA regulatory network in phytoplasma cells during host switching.

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Chlamydia trachomatis σ 28 recognizes the fliC promoter of Escherichia coli and responds to heat shock in chlamydiae

Cited by 21 publications

References 56 publications

Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Selective Promoter Recognition by Chlamydial σ²⁸Holoenzyme

New ex vivo reporter assay system reveals that σ factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants

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Chlamydia trachomatis σ 28 recognizes the fliC promoter of Escherichia coli and responds to heat shock in chlamydiae

Cited by 21 publications

References 56 publications

Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems

Selective Promoter Recognition by Chlamydial σ28Holoenzyme

New ex vivo reporter assay system reveals that σ factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants

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Selective Promoter Recognition by Chlamydial σ²⁸Holoenzyme