Studies On The Cellular Location And Self-interaction Of Rice Ragged Stunt Virus Pns10

Rice ragged stunt virus (RRSV) is the type species of the genus oryzavirus of the family Reoviridae. The genome of RRSV consists of 10 double-stranded RNAs (dsRNAs), encoding eleven proteins including eight structural proteins and three non-structural proteins (Pns6, Pns7 and Pns10) respectively. Rice ragged stunt disease caused by RRSV is a big threat to rice production in some countries of the Southeasten Asia,East Asia and South Asia.In this study, the location of RRSV-Pns10 and its deletion mututions Pns10â–³N and Pns10â–³C in insect cells were identified by direct fluorescence observation and immunofluorescent technique. The results showed that both the intact and the mutant Pns10 with deletions in either N- or C- terminus could form inclusions resembling viroplasm in sf9 cells. The inclusions formed by the N- terminus deleted Pns10 were larger and more dispersed than those formed by the intact Pns10. The inclusions formed by the C- terminus deleted Pns10 showed no differences with those formed by the intact Pns10 in size and distribution.The results of the yeast two-hybrid and co-immunoprecipitation showed that Pns10 can interact with itself and both N- and C-terminal 10 amino acids of Pns10 protein were essential for self-interaction.RRSV-Pns10 located in the cytoplasm, can interact with itself. Bisides, Previously, it has been reported that Pns10 has the activity of ATPase. So Pns10 may be a protein involved in viroplasm formation.