Distinct regions at the N-terminus of the Cucumber necrosis virus coat protein target chloroplasts and mitochondria.

Cucumber necrosis virus (CNV) is a spherical virus consisting of 180 identical coat protein (CP) subunits. The N-terminus of the CP subunit contains a 58aa RNA binding (R) domain and a 34aa arm that connects the R domain to the shell. These regions are known to play critical roles in virus assembly and disassembly. It has recently been shown that a region encompassing the arm can function as a chloroplast transit peptide (TP) in infected plants and that targeting may represent a means for virus particle disassembly. In this study, we further delineate the TP region and show that a 22aa sequence at the N-terminus of the shell enhances chloroplast targeting. We also demonstrate that R domain specifically co-localizes with mitochondria in agroinfiltrated plants. Deletion analyses show that the N-terminal 39 amino acids of the R domain are sufficient for mitochondrial targeting and that this region contains features typical of mitochondrial presequences. The R/arm region is found to be dually targeted to mitochondria and chloroplasts suggesting that this region of the CP plays a critical role in determining the fate of CP during the infection process.