National Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Proc Natl Acad Sci U S A. 2024 Aug 27;121(35):e2403424121. doi: 10.1073/pnas.2403424121. Epub 2024 Aug 19.
Many virus genomes encode proteases that facilitate infection. The molecular mechanism of plant recognition of viral proteases is largely unexplored. Using the system of and cowpea mosaic virus (CPMV), we identified a cowpea lipid transfer protein (LTP1) which interacts with CPMV-encoded 24KPro, a cysteine protease, but not with the enzymatically inactive mutant 24KPro(C166A). Biochemical assays showed that LTP1 inhibited 24KPro proteolytic cleavage of the coat protein precursor large coat protein-small coat protein. Transient overexpression of LTP1 in cowpea reduced CPMV infection, whereas RNA interference-mediated silencing increased CPMV accumulation in cowpea. LTP1 is mainly localized in the apoplast of uninfected plant cells, and after CPMV infection, most of the LTP1 is relocated to intracellular compartments, including chloroplast. Moreover, in stable -transgenic plants, LTP1 repressed soybean mosaic virus (SMV) nuclear inclusion a protease activity, and accumulation of SMV was significantly reduced. We propose that cowpea LTP1 suppresses CPMV and SMV accumulation by directly inhibiting viral cysteine protease activity.
许多病毒基因组编码有助于感染的蛋白酶。植物对病毒蛋白酶识别的分子机制在很大程度上尚未被探索。使用 和豇豆花叶病毒(CPMV)的系统,我们鉴定了一种豇豆脂转移蛋白(LTP1),它与 CPMV 编码的 24KPro(一种半胱氨酸蛋白酶)相互作用,但与无酶活性的突变体 24KPro(C166A)不相互作用。生化分析表明,LTP1 抑制了 24KPro 对半衣壳蛋白前体大衣壳蛋白-小衣壳蛋白的蛋白水解切割。豇豆中 LTP1 的瞬时过表达降低了 CPMV 的感染,而 RNA 干扰介导的 沉默增加了豇豆中 CPMV 的积累。LTP1 主要定位于未感染植物细胞的质外体中,并且在 CPMV 感染后,大部分 LTP1 被重新定位到包括叶绿体在内的细胞内隔室中。此外,在稳定的 -转基因 植物中,LTP1 抑制了大豆花叶病毒(SMV)核包含体 a 蛋白酶活性,并且 SMV 的积累显著减少。我们提出,豇豆 LTP1 通过直接抑制病毒半胱氨酸蛋白酶活性来抑制 CPMV 和 SMV 的积累。