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摘要
The gene encoding a Lon protease homologue has been cloned from Brucella abortus. The putative Brucella abortus Lon shares >60% amino acid identity with its Escherichia coli counterpart and the recombinant form of this protein restores the capacity of an Escherichia coli ion mutant to resist killing by ultraviolet irradiation and regulate the expression of a cpsB::lacZ fusion to wild-type levels. A sigma(32) type promoter was identified upstream of the predicted ion coding region and Northern analysis revealed that transcription of the native Brucella abortus ion increases in response to heat shock and other environmental stresses. ATP-dependent proteolytic activity was also demonstrated for purified recombinant Lon, To evaluate the capacity of the Brucella abortus Lon homologue to function as a stress response protease, the majority of the ion coding region was removed from virulent strain Brucella abortus 2308 via allelic exchange. In contrast to the parent strain, the Brucella abortus ion mutant, designated GR106, was impaired in its capacity to form isolated colonies on solid medium at 41 degrees C and displayed an increased sensitivity to killing by puromycin and H2O2 GR106 also displayed reduced survival in cultured murine macrophages and significant attenuation in BALB/c mice at 1 week post infection compared with the virulent parental strain. Beginning at 2 weeks and continuing for 6 weeks post infection, however, GR106 and 2308 displayed equivalent spleen and liver colonization levels in mice. These findings suggest that the Brucella abortus Lon homologue functions as a stress response protease that is required for wild-type virulence during the initial stages of infection in the mouse model, but is not essential for the establishment and maintenance of chronic infection in this host.
- 出版日期2000-2
