Identification of a quality-control mechanism for mRNA 5′-end capping

Abstract

The 7-methylguanosine cap structure at the 5′-end of eukaryotic mRNAs is a critical determinant of their stability and translational efficiency1–3. It is generally believed that 5’-end capping is a constitutive process that occurs during mRNA maturation and lacks the need for a quality control mechanism to ensure its fidelity. We recently reported that the yeast Rai1 protein has pyrophosphohydrolase activity towards mRNAs lacking a 5’-end cap4. Here we show that, in vitro as well as in yeast cells, Rai1 possess a novel decapping endonuclease activity that can also remove the entire cap structure dinucleotide from an mRNA. Interestingly this activity is targeted preferentially towards mRNAs with unmethylated caps in contrast to the canonical decapping enzyme, Dcp2, that targets mRNAs with a methylated cap. Capped but unmethylated mRNAs generated in yeast cells with a defect in the methyltransferase gene are more stable in a rai1 gene disrupted background. Moreover, rai1Δ yeast cells with wild-type capping enzymes show significant accumulation of mRNAs with 5’-end capping defects under nutritional stress conditions of glucose or amino acid starvation. These findings provide evidence that 5’-end capping is not a constitutive process that necessarily always proceeds to completion and demonstrates that Rai1 plays an essential role in clearing mRNAs with aberrant 5’-end caps. We propose Rai1 is involved in a hitherto-uncharacterized quality control process that ensures mRNA 5’-end integrity by an aberrant-cap mediated mRNA decay mechanism.