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Arabidopsis LSM1 and DCP5 proteins aid evasion of translational repression during Cauliflower Mosaic Virus infection

ABSTRACT: Arabidopsis LSM1 and DCP5 proteins aid evasion of translational repression during Cauliflower Mosaic Virus infection

PROVIDER: PRJNA799378 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17697100_1.fastq.gz	Fastqsanger.gz
	SRR17697100_2.fastq.gz	Fastqsanger.gz
	SRR17697101_1.fastq.gz	Fastqsanger.gz
	SRR17697101_2.fastq.gz	Fastqsanger.gz
	SRR17697102_1.fastq.gz	Fastqsanger.gz

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Arabidopsis LSM1 and DCP5 proteins aid evasion of translational repression during Cauliflower Mosaic Virus infection

Project description:Processing bodies are tightly connected with the RNA silencing machinery. Deletion of processing body components leads to succeptibility of Cauliflower Mosaic Virus to translational repression. Here, we investigated if this succeptibility is mediated through differential sRNA targeting of the viral genome.

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Arabidopsis RNA processing body components LSM1 and DCP5 aid in the evasion of translational repression during Cauliflower mosaic virus infection.

Project description:Viral infections impose extraordinary RNA stress, triggering cellular RNA surveillance pathways such as RNA decapping, nonsense-mediated decay, and RNA silencing. Viruses need to maneuver among these pathways to establish infection and succeed in producing high amounts of viral proteins. Processing bodies (PBs) are integral to RNA triage in eukaryotic cells, with several distinct RNA quality control pathways converging for selective RNA regulation. In this study, we investigated the role of Arabidopsis thaliana PBs during Cauliflower mosaic virus (CaMV) infection. We found that several PB components are co-opted into viral factories that support virus multiplication. This pro-viral role was not associated with RNA decay pathways but instead, we established that PB components are helpers in viral RNA translation. While CaMV is normally resilient to RNA silencing, dysfunctions in PB components expose the virus to this pathway, which is similar to previous observations for transgenes. Transgenes, however, undergo RNA quality control-dependent RNA degradation and transcriptional silencing, whereas CaMV RNA remains stable but becomes translationally repressed through decreased ribosome association, revealing a unique dependence among PBs, RNA silencing, and translational repression. Together, our study shows that PB components are co-opted by the virus to maintain efficient translation, a mechanism not associated with canonical PB functions.

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Project description:Description of gene expression in various types of cauliflower (including Romanesco type) Cauliflower structures develop on inflorescences from plants such as the apetala1 cauliflower double mutant in Arabidopsis thaliana or in the edible Brassica oleracea var botrytis or Romanesco. Whereas the genetic basis for the development of such structure is well understood in the model plant Arabidopsis thaliana, it is less understood in Brassica oleracea. In order to gain insight into the gene expression present in cauliflower and Romanesco curds, we performed RNA-seq analyses of mature cauliflowers and Romanesco curds and from growing cauliflowers curds. This analysis allowed to establish the nature of genes from the APETALA1 and CAULIFLOWER clade that are either mutated or expressed at very low levels in curd structures. These results shed light on the molecular nature of structures that have been selected in the domestication process.

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Project description:Three plants of two species in Brassica genus, i.e. B. rapa cv. "Purple To White Globe" and B. napus cv. "SariGol", highly susceptible and tolerant hosts of Cauliflower mosaic virus (CaMV), respectively, were mechanically inoculated with CaMV strain NY8153.Phosphate buffer was used as control in mock-inoculated plants. RNAs, in three biological repeats, were extracted 20 days post-inoculation (dpi) collecting ten leaves from mock-inoculated and virus-infected plants using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. Five µg of total RNA were resolved in 15% SDS-PAGE gel. Total RNA pattern was visualized upon Ethidium Bromide staining and sRNAs in the size range of 20-30 nt were eluted from gel. sRNAs replicates were quantified and mixed in equal molarity. Libraries were obtained using NEXTflex Small RNA-Seq Kit v3 (Bio Scientific) according to manufacturer’s instructions. Libraries were eluted from a 6% non-denaturing PAGE. The quality of libraries was checked by BioPharma QC, and then, submitted for sequencing with a HiSeq 2500 Illumina platform.

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