Background: Our study investigated the sub-cellular localization of the Parechovirus HPeV 3A nonstructural protein.
Methods: Primers were designed for making wild type HPeV1 and CAV9 2A, 2B, 2C, 2BC and 3A proteins to use for PCR and the pGEM-T Easy vector. The QuikChange II site-directed mutagenesis kit was used to mutate pEGFP-3A using primers. Cells were grown on cover slips and visualized using an Olympus BX41 fluorescence microscope.
Results: We found that EGFP-3A co-localized partially with Golgi and ERGIC. The mutagenesis analysis suggests that the amino acids 39 to 86 of 3A affect distribution and are essential for localization.
Conclusions: Interestingly, co-expression of HPeV 2A with HPeV 3A caused 2A translocation to the nucleolus. The deletion of the TM region in EGFP-2A prevents re-localization.Author(s): Arsalan Salimi, Siamak Amirfakhri