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Chapter 13 : Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants

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Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants, Page 1 of 2

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Abstract:

The bacterial DnaJ-like protein DjlA is composed of two functional domains, an N-terminal transmembrane domain of type III topology and a J domain at the C terminus. The authors investigated the DjlA protein. The plasmid constructs were used to transform and strains. Overexpression of the gene in K-12 resulted in a mucoid phenotype due to colanic acid capsule production. The mutants of all four strains did not exhibit alterations in growth rate in buffered yeast extract (BYE) broth when compared with the appropriate wild type, suggesting that is not an essential gene in . Incubation with a pool of 40% normal human serum revealed that the mutants were as serum-resistant as the parent wild-type strains. The obtained CFU counts for strains R458 and Corby and the corresponding mutants are depicted. Thus, deletion of the gene resulted in reduced virulence in all investigated strains. To our knowledge, this is the first report of an influence on virulence by the gene product in pathogenic bacteria. However, the authors do not know by which mechanism virulence attenuation is achieved in the mutants.

Citation: Brabetz W, Brade H, Frosch M, Lüneberg E. 2002. Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants, p 64-67. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch13

Key Concept Ranking

Bacterial Proteins
0.60302204
Escherichia coli
0.5665815
Legionella pneumophila
0.55664146
0.60302204
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Figures

Image of FIGURE 1
FIGURE 1

(A) Physical map of plasmid pLP028, which harbors the gene from strain Philadelphia (ATCC 33152). (B) Plasmid constructs for overexpression of the gene with its own promoter. (C) Plasmids constructed for overexpression under P control.

Citation: Brabetz W, Brade H, Frosch M, Lüneberg E. 2002. Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants, p 64-67. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch13
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Image of FIGURE 2
FIGURE 2

Infection of HL-60 cells with wild-type and mutant strains. Bacterial CFU were determined at 24, 48, and 72 h postinfection and are shown as means of duplicate values. Experiments were repeated three times.

Citation: Brabetz W, Brade H, Frosch M, Lüneberg E. 2002. Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants, p 64-67. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch13
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References

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1. Blattner, F. R.,, G. Plunkett III,,, C. A. Bloch,, N. T. Perna,, V. Burland,, M. Riley,, J. Collado-Vides,, J. D. Glassner,, C. K. Rode,, G. F. Mayhew,, J. Gregor,, N. W. Davis,, H. A. Kirkpatrick,, M. A. Goeden,, D. J. Rose,, B. Mau,, and Y. Shao. 1997. The complete genome sequence of Escherichia coli K-12. Science 277:14531474.
2. Brabetz, W.,, C. E. Schirmer,, and H. Brade. 2000. 3-Deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferase of Legionella pneumophila transfers two Kdo residues to a structurally different lipid A precursor of Escherichia coli. J. Bacteriol. 182:46544657.
3. Clarke, D. J.,, I. B. Holland,, and A. Jacq. 1997. Point mutations in the transmembrane domain of DjlA, a membrane-linked DnaJ-like protein, abolish its function in promoting colanic acid production via the Res signal transduction pathway. Mol. Microbiol. 25:933944.
4. Clarke, D. J.,, A. Jacq,, and I. B. Holland. 1996. A novel DnaJ-like protein in Escherichia coli inserts into the cytoplasmic membrane with a type III topology. Mol. Microbiol. 20:12731286.
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6. Genevaux, P.,, A. Wawrzynow,, M. Zylicz,, C. Georgopoulos,, and W. L. Kelley. 2001. DjlA is a third DnaK co-chaperone of Escherichia coli and DjlA-mediated induction of colanic acid capsule requires DjlA-DnaK interaction. J. Biol. Chem. 276:79067912.
7. Kelley, W. J.,, and C. Georgopoulos. 1997. Positive control of the two-component RcsC/B signal transduction network by DjlA: a member of the DnaJ family of molecular chaperones in Escherichia coli. Mol. Microbiol. 25:913931.
8. Yura, T.,, H. Mori,, H. Nagai,, T. Nagata,, A. Ishihama,, N. Fujita,, K. Isono,, K. Mizobuchi,, and A. Nakata. 1992. Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region. Nucleic Acids Res. 20:33053308.
9. Zuber, M.,, T. A. Hoover,, and D. L. Court. 1995. Analysis of a Coxiella burnetii gene product that activates capsule synthesis in Escherichia coli: requirement for the heat shock chaperone DnaK and the two-component regulator RcsC. J. Bacterial. 177:42384244.

Tables

Generic image for table
TABLE 1

Overexpression of the gene in and in

Citation: Brabetz W, Brade H, Frosch M, Lüneberg E. 2002. Characterization of the DnaJ-Like Protein DjlA: Virulence Attenuation of Mutants, p 64-67. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch13

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