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Article: Identification of MicroRNA-like RNAs in Mycelial and yeast phases of the thermal dimorphic fungus Penicillium marneffei

TitleIdentification of MicroRNA-like RNAs in Mycelial and yeast phases of the thermal dimorphic fungus Penicillium marneffei
Authors
Issue Date2013
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosntds.org
Citation
PLoS Neglected Tropical Diseases, 2013, v. 7 n. 8, article no. e2398 How to Cite?
AbstractBackground:Penicillium marneffei is the most important thermal dimorphic fungus causing systemic mycosis in China and Southeast Asia. While miRNAs are increasingly recognized for their roles in post-transcriptional regulation of gene expression in animals and plants, miRNAs in fungi were less well studied and their potential roles in fungal dimorphism were largely unknown. Based on P. marneffei genome sequence, we hypothesize that miRNA-like RNAs (milRNAs) may be expressed in the dimorphic fungus.Methodology/Principal Findings:We attempted to identify milRNAs in P. marneffei in both mycelial and yeast phase using high-throughput sequencing technology. Small RNAs were more abundantly expressed in mycelial than yeast phase. Sequence analysis revealed 24 potential milRNA candidates, including 17 candidates in mycelial and seven in yeast phase. Two genes, dcl-1 and dcl-2, encoding putative Dicer-like proteins and the gene, qde-2, encoding Argonaute-like protein, were identified in P. marneffei. Phylogenetic analysis showed that dcl-2 of P. marneffei was more closely related to the homologues in other thermal dimorphic pathogenic fungi than to Penicillium chrysogenum and Aspergillus spp., suggesting the co-evolution of dcl-2 among the thermal dimorphic fungi. Moreover, dcl-2 demonstrated higher mRNA expression levels in mycelial than yeast phase by 7 folds (P<0.001). Northern blot analysis confirmed the expression of two milRNAs, PM-milR-M1 and PM-milR-M2, only in mycelial phase. Using dcl-1KO, dcl-2KO, dclDKO and qde-2KO deletion mutants, we showed that the biogenesis of both milRNAs were dependent on dcl-2 but not dcl-1 or qde-2. The mRNA expression levels of three predicted targets of PM-milR-M1 were upregulated in knockdown strain PM-milR-M1KD, supporting regulatory function of milRNAs.Conclusions/Significance:Our findings provided the first evidence for differential expression of milRNAs in different growth phases of thermal dimorphic fungi and shed light on the evolution of fungal proteins involved in milRNA biogenesis and possible role of post-transcriptional control in governing thermal dimorphism.
Persistent Identifierhttp://hdl.handle.net/10722/191441
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLau, SKPen_US
dc.contributor.authorChow, WNen_US
dc.contributor.authorWong, YPen_US
dc.contributor.authorYeung, MYen_US
dc.contributor.authorBao, Yen_US
dc.contributor.authorZhang, Nen_US
dc.contributor.authorLok, Sen_US
dc.contributor.authorWoo, PCYen_US
dc.contributor.authorYuen, KYen_US
dc.date.accessioned2013-10-15T06:59:34Z-
dc.date.available2013-10-15T06:59:34Z-
dc.date.issued2013en_US
dc.identifier.citationPLoS Neglected Tropical Diseases, 2013, v. 7 n. 8, article no. e2398en_US
dc.identifier.urihttp://hdl.handle.net/10722/191441-
dc.description.abstractBackground:Penicillium marneffei is the most important thermal dimorphic fungus causing systemic mycosis in China and Southeast Asia. While miRNAs are increasingly recognized for their roles in post-transcriptional regulation of gene expression in animals and plants, miRNAs in fungi were less well studied and their potential roles in fungal dimorphism were largely unknown. Based on P. marneffei genome sequence, we hypothesize that miRNA-like RNAs (milRNAs) may be expressed in the dimorphic fungus.Methodology/Principal Findings:We attempted to identify milRNAs in P. marneffei in both mycelial and yeast phase using high-throughput sequencing technology. Small RNAs were more abundantly expressed in mycelial than yeast phase. Sequence analysis revealed 24 potential milRNA candidates, including 17 candidates in mycelial and seven in yeast phase. Two genes, dcl-1 and dcl-2, encoding putative Dicer-like proteins and the gene, qde-2, encoding Argonaute-like protein, were identified in P. marneffei. Phylogenetic analysis showed that dcl-2 of P. marneffei was more closely related to the homologues in other thermal dimorphic pathogenic fungi than to Penicillium chrysogenum and Aspergillus spp., suggesting the co-evolution of dcl-2 among the thermal dimorphic fungi. Moreover, dcl-2 demonstrated higher mRNA expression levels in mycelial than yeast phase by 7 folds (P<0.001). Northern blot analysis confirmed the expression of two milRNAs, PM-milR-M1 and PM-milR-M2, only in mycelial phase. Using dcl-1KO, dcl-2KO, dclDKO and qde-2KO deletion mutants, we showed that the biogenesis of both milRNAs were dependent on dcl-2 but not dcl-1 or qde-2. The mRNA expression levels of three predicted targets of PM-milR-M1 were upregulated in knockdown strain PM-milR-M1KD, supporting regulatory function of milRNAs.Conclusions/Significance:Our findings provided the first evidence for differential expression of milRNAs in different growth phases of thermal dimorphic fungi and shed light on the evolution of fungal proteins involved in milRNA biogenesis and possible role of post-transcriptional control in governing thermal dimorphism.-
dc.languageengen_US
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosntds.org-
dc.relation.ispartofPLoS Neglected Tropical Diseasesen_US
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.titleIdentification of MicroRNA-like RNAs in Mycelial and yeast phases of the thermal dimorphic fungus Penicillium marneffeien_US
dc.typeArticleen_US
dc.identifier.emailLau, SKP: skplau@hkucc.hku.hken_US
dc.identifier.emailChow, WN: chow5810@hku.hken_US
dc.identifier.emailWong, YP: annettew@hku.hken_US
dc.identifier.emailYeung, MY: juyeung@hku.hken_US
dc.identifier.emailBao, Y: baoyj@hku.hken_US
dc.identifier.emailZhang, N: zhangna@hku.hken_US
dc.identifier.emailLok, S: silok@genome.hku.hken_US
dc.identifier.emailWoo, PCY: pcywoo@hkucc.hku.hken_US
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hken_US
dc.identifier.authorityLau, SKP=rp00486en_US
dc.identifier.authorityLok, S=rp00271en_US
dc.identifier.authorityWoo, PCY=rp00430en_US
dc.identifier.authorityYuen, KY=rp00366en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1371/journal.pntd.0002398-
dc.identifier.pmid23991243-
dc.identifier.pmcidPMC3749987-
dc.identifier.scopuseid_2-s2.0-84883319233-
dc.identifier.hkuros225790en_US
dc.identifier.volume7en_US
dc.identifier.spagee2398en_US
dc.identifier.epagee2398en_US
dc.identifier.isiWOS:000323941500055-

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