Disruption of a nonribosomal peptide synthetase in Aspergillus fumigatus eliminates gliotoxin production.
The fungal secondary metabolite gliotoxin produced by Aspergillus fumigatus has been hypothesized to be important in the development of invasive aspergillosis. In this study, we addressed this hypothesis by disrupting a nonribosomal peptide synthetase (NRPS) (encoded by gliP) predicted to be involved in gliotoxin production. Mutants with a disrupted gliP locus failed to produce gliotoxin, which confirmed the role of the NRPS encoded by gliP in gliotoxin biosynthesis. We found no morphological, developmental, or physiological defects in DeltagliP mutant strains. In addition, disruption of gliP resulted in down regulation of gene expression in the gliotoxin biosynthesis gene cluster, which was restored with addition of exogenous gliotoxin. This interesting result suggests a role for gliotoxin in regulating its own production. Culture filtrates from the DeltagliP mutant were unable to inhibit ionomycin-dependent degranulation of mast cells, suggesting a role for gliotoxin in suppressing mast cell degranulation and possibly in disease development. However, the DeltagliP mutant did not have an impact on survival or tissue burden in a murine inhalational model of invasive aspergillosis. This result suggests that gliotoxin is not required for virulence in an immunosuppressed host with an invasive pulmonary infection.
Cramer
Robert A
RA
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA. rcramer@duke.edu
Gamcsik
Michael P
MP
Brooking
Rhea M
RM
Najvar
Laura K
LK
Kirkpatrick
William R
WR
Patterson
Thomas F
TF
Balibar
Carl J
CJ
Graybill
John R
JR
Perfect
John R
JR
Abraham
Soman N
SN
Steinbach
William J
WJ
D000818:Animals; D021461:Animals, Outbred Strains; D001228:Aspergillosis; D001232:Aspergillus fumigatus / Q000378:metabolism* / Q000472:pathogenicity*; D015550:Cell Degranulation; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D015536:Down-Regulation; D016681:Genome, Fungal; D005912:Gliotoxin / Q000096:biosynthesis*; D016867:Immunocompromised Host; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000276:immunology; D051379:Mice; D005810:Multigene Family; D009154:Mutation; D009503:Neutropenia; D010453:Peptide Synthases / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D037521:Virulence Factors
D037521:Virulence Factors; D005912:Gliotoxin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase