A major secreted elastase is essential for pathogenicity of Aeromonas hydrophila.
Aeromonas hydrophila is an opportunistic pathogen and the leading cause of fatal hemorrhagic septicemia in rainbow trout. A gene encoding an elastolytic activity, ahyB, was cloned from Aeromonas hydrophila AG2 into pUC18 and expressed in Escherichia coli and in the nonproteolytic species Aeromonas salmonicida subsp. masoucida. Nucleotide sequence analysis of the ahyB gene revealed an open reading frame of 1,764 nucleotides with coding capacity for a 588-amino-acid protein with a molecular weight of 62,728. The first 13 N-terminal amino acids of the purified protease completely match those deduced from DNA sequence starting at AAG (Lys-184). This finding indicated that AhyB is synthesized as a preproprotein with a 19-amino-acid signal peptide, a 164-amino-acid N-terminal propeptide, and a 405-amino-acid intermediate which is further processed into a mature protease and a C-terminal propeptide. The protease hydrolyzed casein and elastin and showed a high sequence similarity to other metalloproteases, especially with the mature form of the Pseudomonas aeruginosa elastase (52% identity), Helicobacter pylori zinc metalloprotease (61% identity), or proteases from several species of Vibrio (52 to 53% identity). The gene ahyB was insertionally inactivated, and the construct was used to create an isogenic ahyB mutant of A. hydrophila. These first reports of a defined mutation in an extracellular protease of A. hydrophila demonstrate an important role in pathogenesis.
Cascón
A
A
Departamento de Sanidad Animal, Microbiología e Inmunología, Facultad de Veterinaria, Universidad de León, 24071 León, Spain.
Yugueros
J
J
Temprano
A
A
Sánchez
M
M
Hernanz
C
C
Luengo
J M
JM
Naharro
G
G
D016980:Aeromonas hydrophila / Q000472:pathogenicity*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D003227:Conjugation, Genetic; D005393:Fish Diseases / Q000382:microbiology*; D015723:Gene Library; D016905:Gram-Negative Bacterial Infections / Q000662:veterinary*; D007928:Lethal Dose 50; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss; D010196:Pancreatic Elastase / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; C409565:ahyB protein, Aeromonas hydrophila; D010196:Pancreatic Elastase