Hairpin dsRNA does not trigger RNA interference in Candida albicans cells.
RNA interference/silencing mechanisms triggered by double-stranded RNA (dsRNA) have been described in many eukaryotes, including fungi. These mechanisms have in common small RNA molecules (siRNAs or microRNAs) originating from dsRNAs that, together with the effector protein Argonaute, mediate silencing. The genome of the fungal pathogen Candida albicans harbours a well-conserved Argonaute and a non-canonical Dicer, essential members of silencing pathways. Prototypical siRNAs are detected as members of the C. albicans transcriptome, which is potential evidence of RNA interference/silencing pathways in this organism. Surprisingly, expression of a dsRNA a hairpin ADE2 dsRNA molecule to interfere with the endogenous ADE2 mRNA did not result in down-regulation of the message or produce adenine auxotrophic strains. Cell free assays showed that the hairpin dsRNA was a substrate for the putative C. albicans Dicer, discounting the possibility that the nature of the dsRNA trigger affects silencing functionality. Our results suggested that unknown cellular events govern the functionality of siRNAs originating from transgenes in RNA interference/silencing pathways in C. albicans.
Staab
Janet F
JF
Department of Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD, USA. jstaab1@jhmi.edu
White
Theodore C
TC
Marr
Kieren A
KA
D002176:Candida albicans / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D035683:MicroRNAs / Q000235:genetics; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes
D035683:MicroRNAs; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D034741:RNA, Small Interfering