title: Influence of 17β-estradiol on gene expression of Paracoccidioides during mycelia-to-yeast transition.
abstract: BACKGROUND
  Paracoccidioides is the causative agent of paracoccidioidomycosis, a systemic mycosis endemic to Latin America. Infection is initiated by inhalation of conidia (C) or mycelial (M) fragments, which subsequently differentiate into yeast (Y). Epidemiological studies show a striking predominance of paracoccidioidomycosis in adult men compared to premenopausal women. In vitro and in vivo studies suggest that the female hormone (17β-estradiol, E(2)) regulates or inhibits M-or-C-to-Y transition. In this study we have profiled transcript expression to understand the molecular mechanism of how E(2) inhibits M-to-Y transition.
  
  
  METHODS
  We assessed temporal gene expression in strain Pb01 in the presence or absence of E(2) at various time points through 9 days of the M-to-Y transition using an 11,000 element random-shear genomic DNA microarray and verified the results using quantitative real time-PCR. E(2)-regulated clones were sequenced to identify genes and biological function.
  
  
  RESULTS
  E(2)-treatment affected gene expression of 550 array elements, with 331 showing up-regulation and 219 showing down-regulation at one or more time points (p≤0.001). Genes with low expression after 4 or 12 h exposure to E(2) belonged to pathways involved in heat shock response (hsp90 and hsp70), energy metabolism, and several retrotransposable elements. Y-related genes, α-1,3-glucan synthase, mannosyltransferase and Y20, demonstrated low or delayed expression in E(2)-treated cultures. Genes potentially involved in signaling, such as palmitoyltransferase (erf2), small GTPase RhoA, phosphatidylinositol-4-kinase, and protein kinase (serine/threonine) showed low expression in the presence of E(2), whereas a gene encoding for an arrestin domain-containing protein showed high expression. Genes related to ubiquitin-mediated protein degradation, and oxidative stress response genes were up-regulated by E(2).
  
  
  CONCLUSIONS
  This study characterizes the effect of E(2) at the molecular level on the inhibition of the M-to-Y transition and is indicative that the inhibitory actions of E(2) may be working through signaling genes that regulate dimorphism.
authors.affiliation: California Institute for Medical Research, Santa Clara Valley Medical Center, San Jose, California, United States of America.
authors.forename: Jata
authors.initials: J
authors.lastname: Shankar
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Wu
authors.forename: Karl V
authors.initials: KV
authors.lastname: Clemons
authors.forename: Jomar P
authors.initials: JP
authors.lastname: Monteiro
authors.forename: Laurence F
authors.initials: LF
authors.lastname: Mirels
authors.forename: David A
authors.initials: DA
authors.lastname: Stevens
mesh_terms: D000328:Adult; D017931:DNA Primers / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005800:Genes, Fungal / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D025282:Mycelium / Q000187:drug effects*; D010228:Paracoccidioides / Q000187:drug effects* / Q000235:genetics*; D059748:Proteolysis / Q000187:drug effects; D060888:Real-Time Polymerase Chain Reaction; D018626:Retroelements / Q000235:genetics; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013696:Temperature; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D015003:Yeasts / Q000187:drug effects*
chemical_list: D017931:DNA Primers; D018626:Retroelements; D004958:Estradiol