The regulation of leukotoxin production in Actinobacillus actinomycetemcomitans strain JP2.
Actinobacillus actinomycetemcomitans (A.a.) can produce a potent leukotoxin that is thought to be involved in evasion of the host immune response. In order to understand the role of A.a. and its leukotoxin in the initiation and progression of periodontal disease, it is important determine how the expression of A.a. virulence factors might be regulated by the local periodontal micro-environment. To facilitate the measurement of leukotoxin levels, a leukotoxin-beta-galactosidase gene fusion was constructed and recombined into the chromosome of A.a. strain JP2 at the leukotoxin locus. The resulting strain, AAM17, produces beta-galactosidase under control of the leukotoxin promoter. It also produces leukotoxin, since integration of the gene fusion into the chromosome was designed to produce a duplication of the leukotoxin gene. This strain was used to measure the change in leukotoxin level in response to alterations in two environmental signals: iron concentration and oxygen tension. When AAM17 was grown in iron-limited media that did not alter growth rate but did increase the levels of other iron-regulated proteins, the levels of the leukotoxin-beta-galactosidase were similar to those found in AAM17 grown in iron-replete media. These results were confirmed in strains AAM17 and JP2 by leukotoxicity assays and RNA blots. Aerobic growth of AAM17 resulted in a three-fold decrease in leukotoxin beta-galactosidase activity compared with anaerobically grown cells. These results indicate that the A.a. leukotoxin is regulated by some of the environmental signals that may vary in the gingival crevice.
Spitznagel
J
J
Department of Periodontolgy, College of Dentistry, University of Tennessee Center for Health Sciences, Memphis 38163, USA.
Kraig
E
E
Kolodrubetz
D
D
D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics* / Q000378:metabolism* / Q000472:pathogenicity; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D003603:Cytotoxins / Q000096:biosynthesis* / Q000235:genetics; D005098:Exotoxins / Q000096:biosynthesis* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006801:Humans; D007166:Immunosuppressive Agents / Q000378:metabolism*; D007501:Iron / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010510:Periodontal Diseases / Q000382:microbiology; D010957:Plasmids; D015183:Restriction Mapping; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D001427:Bacterial Toxins; D003603:Cytotoxins; D005098:Exotoxins; D007166:Immunosuppressive Agents; C018579:leukotoxin; D007501:Iron; D001616:beta-Galactosidase; D010100:Oxygen