Identification and analysis of genes expressed in the<i>Ustilago maydis</i>dikaryon: uncovering a novel class of pathogenesis genes

Morrison, Erin N.; Donaldson, Michael E.; Saville, Barry J.

doi:10.1080/07060661.2012.697077

Cited by 27 publications

(72 citation statements)

References 54 publications

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“…Analysis of fully sequenced cDNAs (Doyle et al ., ) enabled deletion of a region upstream of as‐um02151 (P as‐um02151 ) in a solo‐pathogenic U. maydis strain (SG200ΔP as‐um02151 ) without removing the 3’ UTR for um02151 , or deleting ncRNA1 , a non‐coding RNA previously described by Morrison et al . () that is antisense to the 3’ UTR of um02150 (Fig. A).…”

Section: Resultsmentioning

confidence: 99%

Ustilago maydis natural antisense transcript expression alters mRNA stability and pathogenesis

Donaldson¹,

Saville

2013

Molecular Microbiology

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Ustilago maydis infection of Zea mays leads to the production of thick-walled diploid teliospores that are the dispersal agent for this pathogen. Transcriptome analyses of this model biotrophic basidiomycete fungus identified natural antisense transcripts (NATs) complementary to 247 open reading frames. The U. maydis NAT cDNAs were fully sequenced and annotated. Strand-specific RT-PCR screens confirmed expression and identified NATs preferentially expressed in the teliospore. Targeted screens revealed four U. maydis NATs that are conserved in a related fungus. Expression of NATs in haploid cells, where they are not naturally occurring, resulted in increased steady-state levels of some complementary mRNAs. The expression of one NAT, as-um02151, in haploid cells resulted in a twofold increase in complementary mRNA levels, the formation of sense–antisense double-stranded RNAs, and unchanged Um02151 protein levels. This led to a model for NAT function in the maintenance and expression of stored teliospore mRNAs. In testing this model by deletion of the regulatory region, it was determined that alteration in NAT expression resulted in decreased pathogenesis in both cob and seedling infections. This annotation and functional analysis supports multiple roles for U. maydis NATs in controlling gene expression and influencing pathogenesis.

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Section: Resultsmentioning

confidence: 99%

Ustilago maydis natural antisense transcript expression alters mRNA stability and pathogenesis

Donaldson¹,

Saville

2013

Molecular Microbiology

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“…(), and total RNA was isolated, precipitated, DNAse I‐treated and screened for gDNA contamination following protocols outlined in Morrison et al . (). RNA quality was assessed through electrophoretic separation of glyoxalated RNA on an agarose gel (1X BPTE), as per Sambrook and Russell ().…”

Section: Methodsmentioning

confidence: 97%

“…Resulting cDNA was diluted eightfold (1:7) with dH 2 O, and RT‐PCR reactions were carried out with an AmpliTaq Gold DNA Polymerase kit following protocols outlined in Morrison et al . (). One third of the RT‐PCR product volume was separated through agarose gel electrophoresis (1X TAE), and was visualized by ethidium bromide staining (0.3 µg ml −1 ; BioShop).…”

Section: Methodsmentioning

confidence: 97%

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Natural antisense transcripts are linked to the modulation of mitochondrial function and teliospore dormancy in Ustilago maydis

Ostrowski

Saville

2017

Molecular Microbiology

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The basidiomycete smut fungus Ustilago maydis causes common smut of corn. This disease is spread through the production of teliospores, which are thick-walled dormant structures characterized by low rates of respiration and metabolism. Teliospores are formed when the fungus grows within the plant, and the morphological steps involved in their formation have been described, but the molecular events leading to dormancy are not known. In U. maydis, natural antisense transcripts (NATs) can function to alter gene expression and many NATs have increased levels in the teliospore. One such NAT is as-ssm1 which is complementary to the gene for the mitochondrial seryl-tRNA synthetase (ssm1), an enzyme important to mitochondrial function. The disruption of ssm1 leads to cell lysis, indicating it is also essential for cellular viability. To assess the function of as-ssm1, it was ectopically expressed in haploid cells, where it is not normally present. This expression led to reductions in growth rate, virulence, mitochondrial membrane potential and oxygen consumption. It also resulted in the formation of as-ssm1/ssm1 double-stranded RNA and increased ssm1 transcript levels, but no change in Ssm1 protein levels was detected. Together, these findings suggest a role for as-ssm1 in facilitating teliospore dormancy through dsRNA formation and reduction of mitochondrial function.

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“…For instance, U. maydis infects and produces corn smut disease only when this fungus has undergone mating and changed its morphology from yeast to dikaryotic hyphae. An intergenic lncRNA transcribed in antisense from its two neighboring genes um02150 and um02151 increases in expression during filamentation and during meiotic spore production and in planta (Morrison et al 2012). Consistently, the loss of this lncRNA significantly reduces the virulence of the fungus.…”

Section: Long Non-coding Rnas In Eukaryotic Genomesmentioning

confidence: 99%

Non-coding RNAs in the development and pathogenesis of eukaryotic microbes

Chacko

Lin

2013

Appl Microbiol Biotechnol

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RNA has long been regarded as the important intermediary in the central dogma of gene expression. Recently, the importance of RNAs in the regulation of gene expression became evident with the identification and characterization of non-protein coding transcripts named noncoding RNA (ncRNA). The ncRNAs, small and long, are ubiquitously present in all three domains of life and are being recognized for their important roles in genome defense and development. Some of the ncRNAs have been associated with diseases and therefore they offer diagnostic and therapeutic potential. In this mini-review, we have highlighted some recent research on the ncRNAs identified in eukaryotic microbes, with special emphasis on fungi that are pathogenic to humans or plants when possible. It is our contention that further elucidation and understanding of ncRNAs will advance our understanding of the development and pathogenesis of eukaryotic microbes and offer alternatives in the diagnosis and treatment of the diseases caused by these pathogens.

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Identification and analysis of genes expressed in theUstilago maydisdikaryon: uncovering a novel class of pathogenesis genes

Cited by 27 publications

References 54 publications

Ustilago maydis natural antisense transcript expression alters mRNA stability and pathogenesis

Ustilago maydis natural antisense transcript expression alters mRNA stability and pathogenesis

Natural antisense transcripts are linked to the modulation of mitochondrial function and teliospore dormancy in Ustilago maydis

Non-coding RNAs in the development and pathogenesis of eukaryotic microbes

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