Proteomic and transcriptomic analysis of Aspergillus fumigatus on exposure to amphotericin B.
Amphotericin B (AMB) is the most widely used polyene antifungal drug for the treatment of systemic fungal infections, including invasive aspergillosis. It has been our aim to understand the molecular targets of AMB in Aspergillus fumigatus by genomic and proteomic approaches. In transcriptomic analysis, a total of 295 genes were found to be differentially expressed (165 upregulated and 130 downregulated), including many involving the ergosterol pathway, cell stress proteins, cell wall proteins, transport proteins, and hypothetical proteins. Proteomic profiles of A. fumigatus alone or A. fumigatus treated with AMB showed differential expression levels for 85 proteins (76 upregulated and 9 downregulated). Forty-eight of them were identified with high confidence and belonged to the above-mentioned categories. Differential expression levels for Rho-GDP dissociation inhibitor (Rho-GDI), secretory-pathway GDI, clathrin, Sec 31 (a subunit of the exocyst complex), and RAB GTPase Ypt51 in response to an antifungal drug are reported here for the first time and may represent a specific response of A. fumigatus to AMB. The expression of some of these genes was validated by real-time reverse transcription-PCR. The AMB responsive genes/proteins observed to be differentially expressed in A. fumigatus may be further explored for novel drug development.
Gautam
Poonam
P
Innate Immunity, National Institute for Research in Reproductive Health, JM Street, Parel, Mumbai, India. taruna_m@hotmail.com
Shankar
Jata
J
Madan
Taruna
T
Sirdeshmukh
Ravi
R
Sundaram
Curam Sreenivasacharlu
CS
Gade
Wasudev Namdeo
WN
Basir
Seemi Farhat
SF
Sarma
Puranam Usha
PU
D000666:Amphotericin B / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D008969:Molecular Sequence Data; D040901:Proteomics; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000187:drug effects / Q000502:physiology
D000935:Antifungal Agents; D005656:Fungal Proteins; D000666:Amphotericin B