A single argonaute gene participates in exogenous and endogenous RNAi and controls cellular functions in the basal fungus Mucor circinelloides.
The mechanism of RNAi is well described in metazoans where it plays a role in diverse cellular functions. However, although different classes of endogenous small RNAs (esRNAs) have been identified in fungi, their biological roles are poorly described due, in part, to the lack of phenotype of mutants affected in the biogenesis of these esRNAs. Argonaute proteins are one of the key components of the RNAi pathways, in which different members of this protein family participate in the biogenesis of a wide repertoire of esRNAs molecules. Here we identified three argonaute genes of the fungus Mucor circinelloides and investigated their participation in exogenous and endogenous RNAi. We found that only one of the ago genes, ago-1, is involved in RNAi during vegetative growth and is required for both transgene-induced RNA silencing and the accumulation of distinct classes of esRNAs derived from exons (ex-siRNAs). Classes I and II ex-siRNAs bind to Ago-1 to control mRNA accumulation of the target protein coding genes. Class III ex-siRNAs do not specifically bind to Ago-1, but requires this protein for their production, revealing the complexity of the biogenesis pathways of ex-siRNAs. We also show that ago-1 is involved in the response to environmental signals, since vegetative development and autolysis induced by nutritional stress are affected in ago-1(-) M. circinelloides mutants. Our results demonstrate that a single Ago protein participates in the production of different classes of esRNAs that are generated through different pathways. They also highlight the role of ex-siRNAs in the regulation of endogenous genes in fungi and expand the range of biological functions modulated by RNAi.
Department of Genetics and Microbiology, Faculty of Biology, University of Murcia, Murcia, Spain.
María
M
Cervantes
Ana
A
Vila
Francisco E
FE
Nicolás
Simon
S
Moxon
Juan P
JP
de Haro
Tamas
T
Dalmay
Santiago
S
Torres-Martínez
Rosa M
RM
Ruiz-Vázquez
D000595:Amino Acid Sequence; D060565:Argonaute Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001329:Autolysis; D015966:Gene Expression Regulation, Fungal; D055786:Gene Knockout Techniques; D005800:Genes, Fungal / Q000235:genetics*; D008969:Molecular Sequence Data; D009089:Mucor / Q000166:cytology* / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012100:Reproduction, Asexual / Q000235:genetics; D013172:Spores, Fungal / Q000502:physiology; D019076:Transgenes / Q000235:genetics
D060565:Argonaute Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering