Functional analysis of the ClpATPase ClpA of Brucella suis, and persistence of a knockout mutant in BALB/c mice The GenBank accession number for the sequence reported in this paper is AJ224881
| Autor: | Stephan Köhler, Jacques Teyssier, Jean-Pierre Liautard, Euloge Ekaza, Laurence Guilloteau |
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| Rok vydání: | 2000 |
| Předmět: |
0303 health sciences
biology 030306 microbiology Intracellular parasite Mutant medicine.disease_cause biology.organism_classification Microbiology Molecular biology 3. Good health BALB/c 03 medical and health sciences Chaperone (protein) medicine biology.protein Brucella suis Heterologous expression Escherichia coli Gene 030304 developmental biology |
| Zdroj: | Microbiology. 146:1605-1616 |
| ISSN: | 1465-2080 1350-0872 |
| DOI: | 10.1099/00221287-146-7-1605 |
| Popis: | The protein ClpA belongs to a diverse group of polypeptides named ClpATPases, which are highly conserved, and which include several molecular chaperones. In this study the gene encoding the 91 kDa protein b-ClpA of the facultative intracellular pathogen Brucella suis, which showed 70% identity to ClpA of Rhodobacter blasticus, was identified and sequenced. Following heterologous expression in Escherichia coli strains SG1126 (ΔclpA) and SG1127 (Δlon ΔclpA), b-ClpA replaced the function of E. coli ClpA, participating in the degradation of abnormal proteins. A b-clpA null mutant of B. suis was constructed, and growth experiments at 37 and 42 °C showed reduced growth rates for the null mutant, especially at the elevated temperature. The mutant complemented by b-clpA and overexpressing the gene was even more impaired at 37 and 42 °C. In intracellular infection of human THP-1 or murine J774 macrophage-like cells, the clpA null mutant and, to a lesser extent, the strain of B. suis overexpressing b-clpA behaved similarly to the wild-type strain. In a murine model of infection, however, the absence of ClpA significantly increased persistence of B. suis. These results showed that in B. suis the highly conserved protein ClpA by itself was dispensable for intramacrophagic growth, but was involved in temperature-dependent growth regulation, and in bacterial clearance from infected BALB/c mice. |
| Databáze: | OpenAIRE |
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