p26 condensates from Pea enation mosaic virus 2 sequester ribosomal components and suppress translation
The p26 movement protein of Pea enation mosaic virus 2 (PEMV2) interacts with the 2'-O-methyltransferase fibrillarin within nuclear and cytoplasmic condensates. This study analyzed global 2'-O-methylation patterns in PEMV2-infected Nicotiana benthamiana using RiboMethSeq, focusing on ribosomal RNAs (rRNAs). The aim was to assess whether PEMV2 disrupts rRNA 2'-O-methylation to interfere with host translation or ribosome assembly. Overall design: To assess whether PEMV2 infection disrupts rRNA 2'-O-methylation, we conducted RiboMethSeq on Mock- and PEMV2-infected plants. Fib2-silenced plants served as a positive control to confirm that Fib2 knockdown reduces 2'-O-methylation. Leaf tissue was harvested 3 days post-infection for total RNA extraction. RNAs were fragmented under alkaline conditions, followed by end-repair and 5' end phosphorylation. Small RNA sequencing libraries were then prepared and sequenced using the Illumina NovaSeq platform with paired-end 100 base pair reads.