title: The ClpP protease is required for the stress tolerance and biofilm formation in Actinobacillus pleuropneumoniae.
abstract: In the respiratory tract and lung tissue, a balanced physiological response is essential for Actinobacillus pleuropneumoniae to survive various types of challenges. ClpP, the catalytic core of the Clp proteolytic complex, is involved in various stresses response and regulation of biofilm formation in many pathogenic bacteria. To investigate the role of ClpP in the virulence of A. pleuropneumoniae, the clpP gene was deleted by homologous recombination, resulting in the mutant strain S8ΔclpP. The reduced growth of S8ΔclpP mutant at high temperatures and under several other stress conditions suggests that the ClpP protein is required for the stress tolerance of A. pleuropneumoniae. Interestingly, we observed that the S8ΔclpP mutant exhibited an increased ability to take up iron in vitro compared to the wild-type strain. We also found that the cells without ClpP displayed rough and irregular surfaces and increased cell volume relative to the wild-type strain using scanning electron microscopy (SEM). Confocal laser scanning microscopy (CLSM) revealed that the S8ΔclpP mutant showed decreased biofilm formation compared to the wild-type strain. We examined the transcriptional profiles of the wild type S8 and the S8ΔclpP mutant strains of A. pleuropneumoniae using RNA sequencing. Our analysis revealed that the expression of 16 genes was changed by the deletion of the clpP gene. The data presented in this study illustrate the important role of ClpP protease in the stress response, iron acquisition, cell morphology and biofilm formation related to A. pleuropneumoniae and further suggest a putative role of ClpP protease in virulence regulation.
authors: State Key Laboratory of Veterinary Biotechnology, Division of Bacterial Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People's Republic of China.
authors: Fang
authors: F
authors: Xie
authors: Yanhe
authors: Y
authors: Zhang
authors: Gang
authors: G
authors: Li
authors: Long
authors: L
authors: Zhou
authors: Siguo
authors: S
authors: Liu
authors: Chunlai
authors: C
authors: Wang
mesh_terms: D016977:Actinobacillus pleuropneumoniae / Q000201:enzymology* / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D000222:Adaptation, Physiological / Q000187:drug effects; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development*; D049071:Endopeptidase Clp / Q000172:deficiency / Q000378:metabolism*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D007501:Iron / Q000378:metabolism / Q000494:pharmacology; D011137:Polystyrenes; D013312:Stress, Physiological / Q000187:drug effects; D013696:Temperature
chemical_list: D011137:Polystyrenes; D007501:Iron; D049071:Endopeptidase Clp