BSCTV C2 Attenuates the Degradation of SAMDC1 to Suppress DNA Methylation-Mediated Gene Silencing in Arabidopsis[W][OA]

This work shows that a geminivirus-encoded silencing suppressor C2 interacts with a plant host cellular component SAMDC1 and attenuates its fast turnover mediated by the 26S proteasome. As a result, it interferes with the host plant’s DNA methylation-related gene silencing mechanism and facilitates geminivirus infection. Plant viruses are excellent tools for studying microbial–plant interactions as well as the complexities of host activities. Our study focuses on the role of C2 encoded by Beet severe curly top virus (BSCTV) in the virus–plant interaction. Using BSCTV C2 as bait in a yeast two-hybrid screen, a C2-interacting protein, S-adenosyl-methionine decarboxylase 1 (SAMDC1), was identified from an Arabidopsis thaliana cDNA library. The interaction was confirmed by an in vitro pull-down assay and a firefly luciferase complemention imaging assay in planta. Biochemical analysis further showed that the degradation of the SAMDC1 protein was inhibited by MG132, a 26S proteasome inhibitor, and that C2 could attenuate the degradation of the SAMDC1 protein. Genetic analysis showed that loss of function of SAMDC1 resulted in reduced susceptibility to BSCTV infection and reduced viral DNA accumulation, similar to the effect of BSCTV C2 deficiency. Bisulfite sequencing analysis further showed that C2 deficiency caused enhanced DNA methylation of the viral genome in infected plants. We also showed that C2 can suppress de novo methylation in the FWA transgenic assay in the C2 transgene background. Overexpression of SAMDC1 can mimic the suppressive activity of C2 against green fluorescent protein–directed silencing. These results suggest that C2 interferes with the host defense mechanism of DNA methylation-mediated gene silencing by attenuating the 26S proteasome-mediated degradation of SAMDC1.

• Publication year

  2011

• Venue

  The Plant Cell

• Publication date

  2011-01-01

• Fields of study

  Biology, Medicine, Environmental Science

• Identifiers
  DOI 10.1105/tpc.110.081695PMID 21245466PMCID PMC3051253
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Overexpression of SAMDC1 can mimic the suppressive activity of C2 against GFP-directed silencing.

  Confidence 0.92

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• BSCTV C2 deficiency increases DNA methylation of the viral genome, and C2 can suppress de novo methylation in the FWA transgenic assay.

  Confidence 0.94

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• Loss of SAMDC1 reduces susceptibility to BSCTV infection and lowers viral DNA accumulation, resembling the effect of BSCTV C2 deficiency.

  Confidence 0.96

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• BSCTV C2 interacts with SAMDC1 and attenuates SAMDC1 degradation by the 26S proteasome.

  Confidence 0.98

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review

• 26s proteasome

  protein complex

  The eukaryotic proteolytic complex responsible for regulated protein degradation in the host cell.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• bsctv c2

  viral protein

  A geminivirus-encoded protein from Beet severe curly top virus that functions as a silencing suppressor in this study.

  Aliases: Beet severe curly top virus C2, C2

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• dna methylation-mediated gene silencing

  gene silencing pathway

  A host gene silencing pathway that uses DNA methylation to suppress gene expression and restrict viral genomes.

  Aliases: DNA methylation-mediated silencing, gene silencing by DNA methylation

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• fwa transgenic assay

  assay

  An Arabidopsis transgene-based assay used to monitor de novo DNA methylation at the FWA locus.

  Aliases: FWA assay

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• gfp-directed silencing

  assay

  A reporter-gene silencing system in which GFP expression is targeted by host silencing mechanisms.

  Aliases: green fluorescent protein-directed silencing, GFP silencing

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• mg132

  chemical compound

  A chemical inhibitor used here to block proteasome-dependent protein degradation.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review
• samdc1

  host protein

  An Arabidopsis thaliana S-adenosyl-methionine decarboxylase 1 protein identified as a C2-interacting host factor.

  Aliases: S-adenosyl-methionine decarboxylase 1

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewB (s683577b42) review

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