An integrated analysis of the effects of Esculentin 1-21 on Saccharomyces cerevisiae.
The antimicrobial peptide esculentin 1-21 (Esc 1-21) is a shorter synthetic version of the 46-residue peptide occurring in the Rana esculenta skin secretion. Here we propose an integrated proteomic and transcriptomic approach to interpret the biological effects of this peptide on Saccharomyces cerevisiae. We further investigated the response to this peptide by correlating the results of the transcriptome and proteome analysis with phenotypic effects. The results show that S. cerevisiae adapts to Esc 1-21 using the High Osmolarity Glycerol (HOG) pathway involved in osmotic tolerance and cell wall maintenance. Comparative proteomics reveals that Esc 1-21 causes downregulation of enzymes of the lower glycolytic pathway and in genes involved in spindle body formation and remodelling of cell-wall synthesis. Moreover the peptide induces downexpression of protein actin within 45 min and cells pre-treated with peptide show less sensitivity to osmotic stress and increased sensitivity to heat shock stress. The results obtained with the two different methodologies are in agreement at the cellular process levels. A combined approach may help elucidate the main aspects related to the effects of this peptide on the eukaryotic cell. The employment of different technologies may reveal the potential and limitations of each adapted approach in a prospective application for drug screening.
Gamberi
Tania
T
Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale G. Morgagni, 50 I-50134 Firenze, Italy.
Cavalieri
Duccio
D
Magherini
Francesca
F
Mangoni
Maria L
ML
De Filippo
Carlotta
C
Borro
Marina
M
Gentile
Giovanna
G
Simmaco
Maurizio
M
Modesti
Alessandra
A
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D003599:Cytoskeleton / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal / Q000235:genetics; D006027:Glycosides / Q000737:chemistry / Q000494:pharmacology*; D006360:Heat-Shock Proteins / Q000235:genetics; D050296:Microbial Viability / Q000187:drug effects; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D009997:Osmotic Pressure / Q000187:drug effects; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D011284:Pregnenolone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics
D000199:Actins; D000935:Antifungal Agents; D006027:Glycosides; D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D029701:Saccharomyces cerevisiae Proteins; C081583:esculentin steroid; D011284:Pregnenolone