Selective capture of transcribed sequences (SCOTS) of Actinobacillus pleuropneumoniae in the chronic stage of disease reveals an HlyX-regulated autotransporter protein

Abstract

Actinobacillus pleuropneumoniae, an important respiratory pathogen in swine, is able to persist in host tissues for extended periods of time. In the study presented here, selective capture of transcribed sequences (SCOTS) analysis was used to identify genes expressed by A. pleuropneumoniae in the chronic stage of the disease (21 days post infection). After isolation and reverse transcription of RNA from infected lungs as well as from culture-grown A. pleuropneumoniae, transcribed A. pleuropneumoniae sequences were captured from infected lung tissue and subjected to a subtractive hybridization procedure of lung-derived against culture-derived A. pleuropneumoniae cDNA. Twenty-nine of the thirty-six genes that were identified as in vivo-expressed are involved in transport or metabolic processes. We identified a surface-associated putative 104 kDa subtilisin-like autotransporter serine protease, designated AasP, which has not been described in A. pleuropneumoniae to date. The gene was shown to be present in all 15 A. pleuropneumoniae serotypes. It is transcribed in porcine lung tissue on days 7 and 21 post infection. Under anaerobic conditions in vitro, its expression depends on the global anaerobic regulator HlyX. To our knowledge, this is the first report of an autotransporter protein being regulated by a global anaerobic regulator.