A genetics screen highlights emerging roles for CPL3, RST1 and URT1 in RNA metabolism and silencing

Li, Ting; Natran, Annelore; Chen, Yanjun; Vercruysse, J.; Wang, Kun; González, Nathalie; Dubois, Marieke

doi:10.1038/s41477-019-0419-7

Cited by 30 publications

(32 citation statements)

References 68 publications

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“…This result confirmed that RST1 functions as a suppressor of S-PTGS comparable with other proteins involved in RNA degradation 25,55–57,59,63 . Of note, the role of RST1 as S-PTGS suppressor is also supported by an independent study published during the reviewing process of our paper 64 .…”

Section: Resultssupporting

confidence: 74%

RST1 and RIPR connect the cytosolic RNA exosome to the Ski complex in Arabidopsis

et al. 2019

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The RNA exosome is a key 3’−5’ exoribonuclease with an evolutionarily conserved structure and function. Its cytosolic functions require the co-factors SKI7 and the Ski complex. Here we demonstrate by co-purification experiments that the ARM-repeat protein RESURRECTION1 (RST1) and RST1 INTERACTING PROTEIN (RIPR) connect the cytosolic Arabidopsis RNA exosome to the Ski complex. rst1 and ripr mutants accumulate RNA quality control siRNAs (rqc-siRNAs) produced by the post-transcriptional gene silencing (PTGS) machinery when mRNA degradation is compromised. The small RNA populations observed in rst1 and ripr mutants are also detected in mutants lacking the RRP45B/CER7 core exosome subunit. Thus, molecular and genetic evidence supports a physical and functional link between RST1, RIPR and the RNA exosome. Our data reveal the existence of additional cytosolic exosome co-factors besides the known Ski subunits. RST1 is not restricted to plants, as homologues with a similar domain architecture but unknown function exist in animals, including humans.

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Section: Resultssupporting

confidence: 74%

RST1 and RIPR connect the cytosolic RNA exosome to the Ski complex in Arabidopsis

et al. 2019

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“…To test the genetic requirements for the semi-lethal effects of MHAL overexpression in an unbiased manner, we mutagenized MHAL overexpressors in the Col-0 background with ethyl methanesulfonate. When we sequenced the genomes of 72 M3 suppressor of MHAL ( somhal ) lines, we found multiple mutations in RST1 and mutations XRN4 , two loci that prevent transgene silencing (Gazzani et al, 2004; Lange et al, 2019; Li et al, 2019) (Table 3) . The most often mutated locus was ACD6 , in 37 of the 72 suppressor lines, with 28 distinct variants (Figure 3D , Table 3) .…”

Section: Resultsmentioning

confidence: 99%

“…Shown are candidate mutations, which are all G>A (C>T) changes, as typical for EMS, from 52 lines in three genes, XRN4, RST1 and ACD6, with multiple hits. Mutations in RST1 and XRN4 are known for transgene silencing (Gazzani et al, 2004;Lange et al, 2019;Li et al, 2019). In the remaining 20 lines, mutations were found, but without any overlapping genes.…”

Section: Injection Of Oocytes and Electrophysiological Measurements mentioning

confidence: 99%

Small proteins modulate ion channel-like ACD6 to regulate immunity inArabidopsis thaliana

Zhu

Neuhäuser

et al. 2021

Preprint

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The quantitative resistance gene ACCELERATED CELL DEATH 6 (ACD6), which encodes a transmembrane protein with intracellular ankyrin repeats, has been implicated in a trade-off between growth and defense among wild strains of Arabidopsis thaliana. Naturally hyperactive alleles of the ACD6-Est-1 type can lead to spontaneous activation of immune responses, although the extent of visible hyperimmunity in strains with this allele varies substantially. We have identified a natural suppressor locus, MODULATOR OF HYPERACTIVE ACD6 1 (MHA1), which codes for a small protein of ~7 kDa that attenuates activity of the ACD6-Est-1 allele. MHA1 and its paralog MHA1-LIKE (MHAL) differentially interact with specific ACD6 variants, and both MHA1 and MHAL peptides can bind to the ACD6 ankyrin repeats. MHAL also enhances accumulation of an ACD6 complex, thereby increasing activity of the ACD6 standard allele. The ACD6 ankyrin repeats are similar to those of transient receptor potential (TRP) ion channels, and several lines of evidence support that increased ACD6 activity is linked to enhanced calcium signaling. Our work highlights how the study of natural variation reveals new aspects of plant immunity.

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“…RST1 is found throughout the multicellular eukaryotic organism, suggesting that RST1 inhibits conserved trafficking components rather than plant-specific factors, and likely a similar molecular breaks function of RST1 is applicable to other multicellular eukaryotic organisms. Recently, two other studies reported that RST1 also plays roles in RNA surveillance pathway, demonstrating that RST1 is a multifunctional protein in plants (Lange et al, 2019;Li et al, 2019b).…”

Section: Discussionmentioning

confidence: 99%

RST1 Is a FREE1 Suppressor That Negatively Regulates Vacuolar Trafficking in Arabidopsis

et al. 2019

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FYVE domain protein required for endosomal sorting1 (FREE1), a plant-specific endosomal sorting complex required for transport-I component, is essential for the biogenesis of multivesicular bodies (MVBs), vacuolar degradation of membrane protein, cargo vacuolar sorting, autophagic degradation, and vacuole biogenesis in Arabidopsis (Arabidopsis thaliana). Here, we report the characterization of RESURRECTION1 (RST1) as a suppressor of free1 that, when mutated as a null mutant, restores the normal MVB and vacuole formation of a FREE1-RNAi knockdown line and consequently allows survival. RST1 encodes an evolutionarily conserved multicellular organism-specific protein, which contains two Domain of Unknown Function 3730 domains, showing no similarity to known proteins, and predominantly localizes in the cytosol. The depletion of FREE1 causes substantial accumulation of RST1, and transgenic Arabidopsis plants overexpressing RST1 display retarded seedling growth with dilated MVBs, and inhibition of endocytosed FM4-64 dye to the tonoplast, suggesting that RST1 has a negative role in vacuolar transport. Consistently, enhanced endocytic degradation of membrane vacuolar cargoes occurs in the rst1 mutant. Further transcriptomic comparison of rst1 with free1 revealed a negative association between gene expression profiles, demonstrating that FREE1 and RST1 have antagonistic functions. Thus, RST1 is a negative regulator controlling membrane protein homeostasis and FREE1mediated functions in plants.

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A genetics screen highlights emerging roles for CPL3, RST1 and URT1 in RNA metabolism and silencing

Cited by 30 publications

References 68 publications

RST1 and RIPR connect the cytosolic RNA exosome to the Ski complex in Arabidopsis

RST1 and RIPR connect the cytosolic RNA exosome to the Ski complex in Arabidopsis

Small proteins modulate ion channel-like ACD6 to regulate immunity inArabidopsis thaliana

RST1 Is a FREE1 Suppressor That Negatively Regulates Vacuolar Trafficking in Arabidopsis

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