Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway

A hot pepper (Capsicum annuum) cDNA clone encoding pathogenesis-related protein 10 (CaPR-10) was isolated by differential screening of a cDNA library prepared from pepper leaves inoculated with tobacco mosaic virus pathotype (TMV-P₀). CaPR-10 transcripts were induced in the incompatible interaction with TMV-P₀ or Xanthomonas campestris pv. vesicatoria (Xcv) but not induced in the compatible interaction. Characterization of enzymatic properties of CaPR-10 indicated that the recombinant protein exhibits a ribonucleolytic activity against TMV RNA, as well as against pepper total RNA, and shows its putative antiviral activity in several conditions. The CaPR-10 protein existed at very low level in leaf tissue but was dramatically induced as soon as plants were inoculated with TMV-P₀, and this was correlated with the increase of its ribonucleolytic activity. Immunoblot analysis and pull-down assays using proteins extracted from pepper leaves showed that TMV-P₀ inoculation led to the phosphorylation of CaPR-10, a modification that should affect its capacity for RNase function. We present data that the induction and subsequent phosphorylation of CaPR-10 increased its ribonucleolytic activity to cleave invading viral RNAs, and this activity should be important to its antiviral pathway during viral attack in vivo.