Abstract
As a core subunit of the SCF complex that promotes protein degradation through the 26S proteasome, S-phase kinase-associated protein 1 (SKP1) plays important roles in multiple cellular processes in eukaryotes, including gibberellin (GA), jasmonate, ethylene, auxin and light responses. P7-2 encoded by Rice black streaked dwarf virus (RBSDV), a devastating viral pathogen that causes severe symptoms in infected plants, interacts with SKP1 from different plants. However, whether RBSDV P7-2 forms a SCF complex and targets host proteins is poorly understood. In this study, we conducted yeast two-hybrid assays to further explore the interactions between P7-2 and 25 type I Oryza sativa SKP1-like (OSK) proteins, and found that P7-2 interacted with eight OSK members with different binding affinity. Co-immunoprecipitation assay further confirmed the interaction of P7-2 with OSK1, OSK5 and OSK20. It was also shown that P7-2, together with OSK1 and O. sativa Cullin-1, was able to form the SCF complex. Moreover, yeast two-hybrid assays revealed that P7-2 interacted with gibberellin insensitive dwarf2 (GID2) from rice and maize plants, which is essential for regulating the GA signaling pathway. It was further demonstrated that the N-terminal region of P7-2 was necessary for the interaction with GID2. Overall, these results indicated that P7-2 functioned as a component of the SCF complex in rice, and interaction of P7-2 with GID2 implied possible roles of the GA signaling pathway during RBSDV infection.
Highlights
The S-phase kinase-associated protein 1 (SKP1) is a core component of the SKP1/Cullin/Fbox (SCF) complex, an E3 ubiquitin ligase that triggers protein degradation through the 26S proteasome [1,2]
P7-2 was fused to the GAL4 DNA-binding domain (BD), and the 25 Oryza sativa SKP1-like (OSK) were fused to the GAL4 activation domain (AD), respectively
Only the diploid yeast strains produced from mating of BD-P7-2 in Y187 and AD-OSKs (OSK1, OSK4, OSK5, OSK6, OSK9, OSK10, OSK20 and OSK25) in AH109, respectively, were able to grow on selective media (SD/-Leu/Trp/-Ade/-His) that contained X-α-gal (Fig 1)
Summary
The S-phase kinase-associated protein 1 (SKP1) is a core component of the SKP1/Cullin/Fbox (SCF) complex, an E3 ubiquitin ligase that triggers protein degradation through the 26S proteasome [1,2]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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