The transcriptome analysis of early morphogenesis in Paracoccidioides brasiliensis mycelium reveals novel and induced genes potentially associated to the dimorphic process.
BACKGROUND
Paracoccidioides brasiliensis is a human pathogen with a broad distribution in Latin America. The fungus is thermally dimorphic with two distinct forms corresponding to completely different lifestyles. Upon elevation of the temperature to that of the mammalian body, the fungus adopts a yeast-like form that is exclusively associated with its pathogenic lifestyle. We describe expressed sequence tags (ESTs) analysis to assess the expression profile of the mycelium to yeast transition. To identify P. brasiliensis differentially expressed sequences during conversion we performed a large-scale comparative analysis between P. brasiliensis ESTs identified in the transition transcriptome and databases.


RESULTS
Our analysis was based on 1107 ESTs from a transition cDNA library of P. brasiliensis. A total of 639 consensus sequences were assembled. Genes of primary metabolism, energy, protein synthesis and fate, cellular transport, biogenesis of cellular components were represented in the transition cDNA library. A considerable number of genes (7.51%) had not been previously reported for P. brasiliensis in public databases. Gene expression analysis using in silico EST subtraction revealed that numerous genes were more expressed during the transition phase when compared to the mycelial ESTs 1. Classes of differentially expressed sequences were selected for further analysis including: genes related to the synthesis/remodeling of the cell wall/membrane. Thirty four genes from this family were induced. Ten genes related to signal transduction were increased. Twelve genes encoding putative virulence factors manifested increased expression. The in silico approach was validated by northern blot and semi-quantitative RT-PCR.


CONCLUSIONS
The developmental program of P. brasiliensis is characterized by significant differential positive modulation of the cell wall/membrane related transcripts, and signal transduction proteins, suggesting the related processes important contributors to dimorphism. Also, putative virulence factors are more expressed in the transition process suggesting adaptation to the host of the yeast incoming parasitic phase. Those genes provide ideal candidates for further studies directed at understanding fungal morphogenesis and its regulation.
Bastos
Karinne P
KP
Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, Goiás, Brasil. karinnebastos@yahoo.com.br <karinnebastos@yahoo.com.br>
Bailão
Alexandre M
AM
Borges
Clayton L
CL
Faria
Fabricia P
FP
Felipe
Maria S S
MS
Silva
Mirelle G
MG
Martins
Wellington S
WS
Fiúza
Rogério B
RB
Pereira
Maristela
M
Soares
Célia M A
CM
D000222:Adaptation, Physiological / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002473:Cell Wall / Q000378:metabolism / Q000648:ultrastructure; D019295:Computational Biology; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics; D025282:Mycelium / Q000235:genetics / Q000254:growth & development*; D010228:Paracoccidioides / Q000235:genetics / Q000254:growth & development*; D012331:RNA, Fungal / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000235:genetics; D014158:Transcription, Genetic
D004271:DNA, Fungal; D012331:RNA, Fungal