Functional characterization of the Aspergillus fumigatus calcineurin.
Aspergillus fumigatus is an aggressive opportunistic pathogen of humans as well as a major allergen. Environmental sensing and retrieving essential nutrients from the environment are general metabolic traits associated with the growth of this saprophytic fungus. Two important mediators of calcium signals in eukaryotic cells are the Ca(2+)-binding protein calmodulin and the Ca(2+)/calmodulin-dependent phosphatase calcineurin. Calcineurin is a heterodimer that consists of a catalytic subunit A and a Ca(2+)/calmodulin binding unit. We deleted the A. fumigatus calA gene, which encodes the calcineurin A catalytic subunit, and demonstrated that this gene is not essential in this fungus. The DeltacalA mutant strain has severe defects in growth extension, branching and conidial architecture. Furthermore, the A. fumigatus DeltacalA mutant strain has decreased fitness in a low dose murine infection and cannot grow in fetal bovine serum (FBS). After potassium phosphate was added to liquid FBS, the DeltacalA mutant strain could grow with the characteristic phenotype of the DeltacalA mutation. When A. fumigatus calcineurin is inhibited by tacrolimus in a phosphate depleted medium, there is a reduction in the inorganic phosphate transport and six putative phosphate transporter genes have altered mRNA levels. However, there is no effect on the acid phosphatase activity. These results suggest that calcineurin is involved in the regulation of the PHO pathway in A. fumigatus. Our work on calcineurin opens new venues for the research on sensing and nutrient acquisition in A. fumigatus.
da Silva Ferreira
Márcia Eliana
ME
Faculdade de Ciências Farmacêuticas de Ribeirão, Universidade de São Paulo, Brazil.
Heinekamp
Thorsten
T
Härtl
Albert
A
Brakhage
Axel A
AA
Semighini
Camile P
CP
Harris
Steven D
SD
Savoldi
Marcela
M
de Gouvêa
Paula Fagundes
PF
de Souza Goldman
Maria Helena
MH
Goldman
Gustavo H
GH
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D019703:Calcineurin / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain / Q000235:genetics; D002417:Cattle; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D005312:Fetal Blood / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D025301:Hyphae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D026901:Membrane Transport Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D009154:Mutation; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013172:Spores, Fungal / Q000235:genetics / Q000648:ultrastructure; D016559:Tacrolimus / Q000494:pharmacology; D013997:Time Factors
D003470:Culture Media; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D010710:Phosphates; D012333:RNA, Messenger; D019703:Calcineurin; D000135:Acid Phosphatase; D016559:Tacrolimus