Translatome profiling of Drosophila intestinal enterocytes upon Dietary Restriction

We determined the actively translated mRNA induced/repressed by a Dietary Restriction intervention specifically in the differentiated enterocytes of the intestine. For this, we did a "genetic dissection" by expressing a tagged ribosomal subunit (RpL13A-His6FLAG) under the control of the tissue specific Myo1A-GAL4 driver, targetting its expression exclusively to the intestinal enterocytes. By performing a anti-flag pulldown we are then able to enrich our mRNA pool for transcripts that are directly bound to ribosomes and being actively translated. In this manner, we become one step closer to protein synthesis while profiling gene expression. Overall design: 2-3 day old Myo1A-GS X Ribotag Drosophila adult flies were placed on Ad Libitum (AL) or Dietary Restriction (DR) diets and maintained for 10 days prior to polysomal pulldown.