Identifying tunicamycin targets on three microarray generations. Barcode intensity data are normalized according to a DMSO reference treatment. Blue dots represent non-essential genes, red dots represent essential genes and grey dots are genes that are not annotated. Log2 ratios are calculated as a measure of change in barcode intensity (vertical axis) across all genes (horizontal axis). Ratios below 0 have been removed for clarity. Log2 scales differ based on optimal dynamic range between baseline and ALG7. Higher ratios correspond to greater abundance of barcode from reference to treatment. In all three analyses, ALG7 was correctly identified as the primary target of tunicamycin. Several additional genes previously determined to be resistant to tunicamycin, were most discernibly identified in the tiling data, but less so using TAG4 (the current microarray standard) and TAG3. These include ADO1, BCK1, FYV8, GET2, HAC1 and IRE1. Furthermore, the genes COP1 and RER2, known to be involved in ER to Golgi vesicle-mediated transport, showed up as sensitive to tunicamycin in our screen.
Ammar et al. BMC Genomics 2009 10:471 doi:10.1186/1471-2164-10-471