Transcriptional and polysomal profile of poliovirus-infected HUVECs

Translation initiation of eukaryotic mRNAs mostly occurs by the cap-dependent ribosome scanning mechanism. However, certain mRNAs are translated by ribosome assembly at internal ribosome entry sites (IRES), a mechanism that allows the synthesis of certain proteins when cap-dependent translation is inhibited by cellular stress. Whether IRES-mediated translation occurs in stressed human endothelial cells (EC) is unknown. We performed whole genome microarray analysis of polyribosomal mRNA (43,203 sequences) from virus-infected EC to identify IRES-containing mRNAs. RNA from polysome fractions or total RNA were isolated from control or PV-infected HUVECs using Trizol LS (Invitrogen, Carlsbad, CA) and then subjected to the whole human genome-wide microarray analysis (43,203 transcripts, Agilent, Santa Clara, CA) to obtain the translational profile.