Two classes of small antisense RNAs in fungal RNA silencing triggered by non-integrative transgenes

Nicolás, Francisco E.; Torres‐Martínez, Santiago; Ruiz‐Vázquez, Rosa M.

doi:10.1093/emboj/cdg384

Cited by 132 publications

(193 citation statements)

References 38 publications

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“…RNA gel blot was carried out as described previously (Helber et al, 2011). The analysis of siRNAs derived from GigmPT was performed as previously described ( Nicolás et al, 2003 andPall andHamilton, 2008).…”

Section: Host-induced Gene Silencingmentioning

confidence: 99%

Arbuscular Mycorrhizal Symbiosis Requires a Phosphate Transceptor in the Gigaspora margarita Fungal Symbiont

et al. 2016

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The definitive version is available at: La versione definitiva è disponibile alla URL: [http://www.sciencedirect.com AbstractThe majority of terrestrial vascular plants are capable of forming mutualistic associations with obligate biotrophic arbuscular mycorrhizal (AM) fungi from the phylum Glomeromycota. This mutualistic symbiosis provides carbohydrates to the fungus, and reciprocally improves plant phosphate uptake. AM fungal transporters can acquire phosphate from the soil through the hyphal networks. Nevertheless, the precise functions of AM fungal phosphate transporters, and whether they act as sensors or as nutrient transporters, in fungal signal transduction remain unclear. Here, we report a high-affinity phosphate transporter GigmPT from Gigaspora margarita that is required for AM symbiosis. Host-induced gene silencing of GigmPT hampers the development of G. margarita during AM symbiosis. Most importantly, GigmPT functions as a phosphate transceptor in G. margarita regarding the activation of the phosphate signaling pathway as well as the protein kinase A signaling cascade. Using the substituted-cysteine accessibility method, we identified residues A146 (in transmembrane domain [TMD] IV) and Val357 (in TMD VIII) of GigmPT, both of which are critical for phosphate signaling and transport in yeast during growth induction. Collectively, our results provide significant insights into the molecular functions of a phosphate transceptor from the AM fungus G. margarita.

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Section: Host-induced Gene Silencingmentioning

confidence: 99%

Arbuscular Mycorrhizal Symbiosis Requires a Phosphate Transceptor in the Gigaspora margarita Fungal Symbiont

et al. 2016

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“…On the one hand, rearrangements may affect copy number determinations (even using Southern blotting techniques, which frequently miss smaller variations), and on the other hand, agarose gel analysis of amplified segments of genomic DNA is insufficient to provide reliable quantification. Nevertheless, combined analysis of these two methods may be used: Fr example, copy numbers of pXLNRir do not correlate with a higher degree of silencing efficiency, a situation similar to others reported (Nakayashiki et al 2005;Nicolas et al 2003;Tanguay et al 2006;Fitzgerald et al 2004;Henry et al 2007;Yamada et al 2007;Walti et al 2006). Apart from genome instability and the occurrence of rearrangements, other factors may be responsible for a lower silencing efficiency of integrated hairpin-expression units.…”

Section: Discussionmentioning

confidence: 67%

Efficient cloning system for construction of gene silencing vectors in Aspergillus niger

Oliveira

Veen

Graaff

2008

Appl Microbiol Biotechnol

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An approach based on Gateway recombination technology to efficiently construct silencing vectors was developed for use in the biotechnologically important fungus Aspergillus niger. The transcription activator of xylanolytic and cellulolytic genes XlnR of A. niger was chosen as target for gene silencing. Silencing was based on the expression vector pXLNRir that was constructed and used in co-transformation. From all the strains isolated (N=77), nine showed poor xylan-degrading activities in two semiquantitative plate assays testing different activities for xylan degradation. Upon induction on D-xylose, transcript levels of xlnR were decreased in the xlnR-silenced strains, compared to a wild-type background. Under these conditions, the transcript levels of xyrA and xynB (two genes regulated by XlnR) were also decreased for these xlnRsilenced strains. These results indicate that the newly developed system for rapid generation of silencing vectors is an effective tool for A. niger, and this can be used to generate strains with a tailored spectrum of enzyme activities or product formation by silencing specific genes encoding, e.g., regulators such as XlnR.

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“…The pH was adjusted to 4.5 and 3.2 for mycelial and colonial growth, respectively. Transformation was carried out as described previously (21). Illumination conditions were as previously described (17).…”

Section: Methodsmentioning

confidence: 99%

Non-AUG Translation Initiation of a Fungal RING Finger Repressor Involved in Photocarotenogenesis

Murcia

Lorca-Pascual

Garre

et al. 2007

Journal of Biological Chemistry

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The RING finger protein CrgA acts as a negative regulator of light-induced carotene biosynthesis in the fungus Mucor circinelloides. Sequence analysis of the crgA coding region upstream of the first AUG codon revealed the existence of an additional non-canonical RING finger domain at the most N-terminal end of the protein. The newly identified RING finger domain is required for CrgA to regulate photocarotenogenesis, as deduced from site-directed mutagenesis experiments. The role of both RING finger domains in the stability of CrgA has been investigated in a yeast system. Wild type CrgA, but not the RING finger deleted forms, is highly unstable and is stabilized by inhibition of the proteasome function, which suggests that native CrgA is degraded by the proteasome and that active RING finger domains are required for proteasome-mediated CrgA degradation. To identify the translation start of CrgA, a mutational analysis of putative initiation codons in the 5 region of the crgA gene was accomplished. We demonstrated that a GUG codon located upstream of the first AUG is the sole initiator of CrgA translation. To our knowledge, this is the first report of a naturally occurring non-AUG start codon for a RING finger regulatory protein. A combination of suboptimal translation initiation and proteasome degradation may help to maintain the low cellular levels of CrgA observed in wild type cells, which is probably required for accurate regulation of photocarotenogenesis.

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Two classes of small antisense RNAs in fungal RNA silencing triggered by non-integrative transgenes

Cited by 132 publications

References 38 publications

Arbuscular Mycorrhizal Symbiosis Requires a Phosphate Transceptor in the Gigaspora margarita Fungal Symbiont

Arbuscular Mycorrhizal Symbiosis Requires a Phosphate Transceptor in the Gigaspora margarita Fungal Symbiont

Efficient cloning system for construction of gene silencing vectors in Aspergillus niger

Non-AUG Translation Initiation of a Fungal RING Finger Repressor Involved in Photocarotenogenesis

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