title: Efficient translation of synthetic and natural mRNAs in an mRNA-dependent cell-free system from the dimorphic fungus Candida albicans.
abstract: An mRNA-dependent cell-free translation system has been developed from the human pathogenic fungus Candida albicans using either S30 or S100 lysates prepared from glass-bead-disrupted whole cells. Translation of the synthetic template poly(U) in this system is highly efficient at temperatures up to 37 degrees C and is ATP-dependent. Studies using a range of elongation-specific inhibitors suggest that the mechanism of translational elongation in C. albicans is similar to that of another yeast, Saccharomyces cerevisiae. A micrococcal-nuclease-treated C. albicans S100 lysate was able to translate exogenously-supplied homologous mRNAs, and a range of heterologous natural mRNAs, using an initiation mechanism that is inhibited by the antibiotic edeine and the 5' cap analogue 7-methylguanosine 5'-monophosphate (m7GMP). As with cell-free lysates prepared from S. cerevisiae, the C. albicans lysate is unable to initiate translation upon natural mRNAs at temperatures above 20 degrees C.
authors.affiliation: Biological Laboratory, University of Kent, Canterbury, UK.
authors.forename: D R
authors.initials: DR
authors.lastname: Colthurst
authors.forename: P
authors.initials: P
authors.lastname: Chalk
authors.forename: M
authors.initials: M
authors.lastname: Hayes
authors.forename: M F
authors.initials: MF
authors.lastname: Tuite
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000935:Antifungal Agents / Q000494:pharmacology; D002176:Candida albicans / Q000235:genetics*; D008836:Micrococcal Nuclease / Q000378:metabolism; D010441:Peptide Chain Elongation, Translational; D010442:Peptide Chain Initiation, Translational; D011072:Poly U / Q000378:metabolism; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013696:Temperature; D013698:Templates, Genetic
chemical_list: D000935:Antifungal Agents; D012331:RNA, Fungal; D012333:RNA, Messenger; D011072:Poly U; D000255:Adenosine Triphosphate; D008836:Micrococcal Nuclease