Insights into the relation between mrna and protein expression patterns: ii. Experimental observations in <i>Escherichia coli</i>1

Lee, Pat S.; Shaw, Leah B.; Choe, Leila H.; Mehra, Amit; Hatzimanikatis, Vassily; Lee, Kelvin H.

doi:10.1002/bit.10841

Cited by 75 publications

(56 citation statements)

References 19 publications

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“…In particular, promoter activities as measured in this study do not always correlate with protein abundance because of posttranscriptional regulation and protein turnover (41). In vivo protein quantification is possible using epitope tagging (14), but this demanding technique is difficult to apply to global screening.…”

Section: Discussionmentioning

confidence: 89%

Antigen selection based on expression levels during infection facilitates vaccine development for an intracellular pathogen

Rollenhagen

Sörensen

Rizos

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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Vaccines effective against intracellular pathogens could save the lives of millions of people every year, but vaccine development has been hampered by the slow largely empirical search for protective antigens. In vivo highly expressed antigens might represent a small attractive antigen subset that could be rapidly evaluated, but experimental evidence supporting this rationale, as well as practical strategies for its application, is largely lacking because of technical difficulties. Here, we used Salmonella strains expressing differential amounts of a fluorescent model antigen during infection to show that, in a mouse typhoid fever model, CD4 T cells preferentially recognize abundant Salmonella antigens. To identify a large number of natural Salmonella antigens with high expression levels during infection, we used a quantitative in vivo screening strategy. Immunization studies with five particularly attractive candidates revealed two highly protective antigens that might permit the development of an improved typhoid fever vaccine. In conclusion, we have established a rationale and an experimental strategy that will substantially facilitate vaccine development for Salmonella and possibly other intracellular pathogens.

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Section: Discussionmentioning

confidence: 89%

Antigen selection based on expression levels during infection facilitates vaccine development for an intracellular pathogen

Rollenhagen

Sörensen

Rizos

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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“…In yeast, GLX1 knockouts are unaffected by osmotic and heat stress, indicating that redundant systems must be able to compensate for the absence of GLX1 during exposure to these stress conditions (30). The relationship between mRNA and protein expression has been investigated in several recent large-scale studies (28,32,43). These studies show that mRNA expression levels cannot be consistently relied on as a predictor of protein abundance (since cells can control the abundance of individual proteins at the transcriptional or translational level).…”

Section: Discussionmentioning

confidence: 99%

Dehydration-Specific Induction of Hydrophilic Protein Genes in the Anhydrobiotic Nematode Aphelenchus avenae

et al. 2004

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Some organisms can survive exposure to extreme desiccation by entering a state of suspended animation known as anhydrobiosis. The free-living nematode Aphelenchus avenae can be induced to enter the anhydrobiotic state by exposure to a moderate reduction in relative humidity. During this preconditioning period, the nematode accumulates large amounts of the disaccharide trehalose, which is thought to be necessary, but not sufficient, for successful anhydrobiosis. To identify other adaptations that are required for anhydrobiosis, we developed a novel SL1-based mRNA differential display technique to clone genes that are upregulated by dehydration in A. avenae. Three such genes, Aav-lea-1, Aav-ahn-1, and Aav-glx-1, encode, respectively, a late embryogenesis abundant (LEA) group 3 protein, a novel protein that we named anhydrin, and the antioxidant enzyme glutaredoxin. Strikingly, the predicted LEA and anhydrin proteins are highly hydrophilic and lack significant secondary structure in the hydrated state. The dehydration-induced upregulation of Aav-lea-1 and Aav-ahn-1 was confirmed by Northern hybridization and quantitative PCR experiments. Both genes were also upregulated by an osmotic upshift, but not by cold, heat, or oxidative stress. Experiments to investigate the relationship between mRNA levels and protein expression for these genes are in progress. LEA proteins occur commonly in plants, accumulating during seed maturation and desiccation stress; the presence of a gene encoding an LEA protein in an anhydrobiotic nematode suggests that some mechanisms of coping with water loss are conserved between plants and animals.

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“…Since it has been clearly shown that mRNA levels do not always accurately predict the amount of protein synthesized from a given gene (e.g. Lee et al, 2003), future experiments including both mRNA and protein expression measurements will be necessary to elucidate the contributions of s B -dependent gene expression to expressed protein levels.…”

Section: B -Dependent Genes and Gene Expression In Virulencementioning

confidence: 99%

σ B-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment

et al. 2004

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Listeria monocytogenes must overcome a variety of stress conditions in the host digestive tract to cause foodborne infections. The alternative sigma factor s B , encoded by sigB, is responsible for regulating transcription of several L. monocytogenes virulence and stress-response genes, including genes that contribute to establishment of gastrointestinal infections. A quantitative RT-PCR assay was used to measure mRNA transcript accumulation for the virulence genes inlA and bsh, the stress-response genes opuCA and lmo0669 (encoding a carnitine transporter and an oxidoreductase, respectively) and the housekeeping gene rpoB. Assays were conducted on mid-exponential phase L. monocytogenes cells exposed to conditions reflecting osmotic (0?3 M NaCl) or acid (pH 4?5) conditions typical for the human intestinal lumen. In exponential-phase cells, as well as under osmotic and acid stress, inlA, opuCA and bsh showed significantly lower absolute expression levels in a L. monocytogenes DsigB null mutant compared to wild-type. A statistical model that normalized target gene expression relative to rpoB showed that accumulation of inlA, opuCA and bsh transcripts was significantly increased in the wild-type strain within 5 min of acid and osmotic stress exposure; lmo0669 transcript accumulation increased significantly only after acid exposure. It was concluded that s B is essential for rapid induction of the tested stress-response and virulence genes under conditions typically encountered during gastrointestinal passage. As inlA, bsh and opuCA are critical for gastrointestinal infections in animal models, the data also suggest that s B contributes to the ability of L. monocytogenes to cause foodborne infections.

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Insights into the relation between mrna and protein expression patterns: ii. Experimental observations in Escherichia coli1

Cited by 75 publications

References 19 publications

Antigen selection based on expression levels during infection facilitates vaccine development for an intracellular pathogen

Antigen selection based on expression levels during infection facilitates vaccine development for an intracellular pathogen

Dehydration-Specific Induction of Hydrophilic Protein Genes in the Anhydrobiotic Nematode Aphelenchus avenae

σ B-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment

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