Project description:We performed immunoprecipitation analysis with leaves from 35S::NusG:MYC transgenic lines throung the MYCantibody, and the product was subjected to LC-MS analysis.
Project description:The mechanistic connection between the THO/TREX complex and RdDM remains poorly understood. To address this, we performed EMS mutagenesis in a transgenic Col-0 background harboring pro2×35S::LUC and pro2×35S::NPTII reporters and homozygous for rdr6, and identified a low-fluorescence mutant, tho2-8. Unlike null alleles, tho2-8 is a viable homozygous allele, offering a unique opportunity to explore THO2 function in DNA methylation. Here, we report that THO2 is required for proper siRNA accumulation and RdDM activity, revealing a previously uncharacterized role for the THO/TREX complex in epigenetic gene silencing. This study expands our understanding of the molecular coordination between RNA metabolism and epigenetic regulation in plants.
Project description:To identify possible mitochondrial DNA binding targets of SHOT1/MTERF18 (AT3G60400), C-terminal GFP fusion constructs under either a constitutive 35S promoter (35S::SHOT1-GFP) or the SHOT1 native promoter (SHOT1p::SHOT1-GFP) were made. The transgenes were introduced into the shot1-2 mutant background (Kim et al., 2012, Plant Cell) and verified to be functional based on recovery of shot1 from growth defects. As a control, a transgenic line harboring mitochondria-targeted GFP (Mito-GFP) under the 35S promoter was used. All enriched peaks in SHOT1-GFP samples are located upstream of different tRNA genes: trnS, trnM, trnG and trnF
