1887

Abstract

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 , which showed 70% identity to ClpA of , was identified and sequenced. Following heterologous expression in strains SG1126 (Δ) and SG1127 (Δ Δ), b-ClpA replaced the function of ClpA, participating in the degradation of abnormal proteins. A null mutant of 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 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 null mutant and, to a lesser extent, the strain of overexpressing behaved similarly to the wild-type strain. In a murine model of infection, however, the absence of ClpA significantly increased persistence of . These results showed that in 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.

Keyword(s): Brucella , chaperone , ClpA and ClpATPase
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2000-07-01
2020-04-02
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