The regulation mechanism of HLA class II gene expression at the level of mRNA stability

The number of major histocompatibility complex (MHC) class II antigens may be regulated at different levels. Although transcriptional regulation has been studied most intensely, evidence for control mechanisms acting on the stability of MHC class II mRNAs has been reported. We have previously shown, in fact, that the half-life of MHC class II mRNA rapidly decreases in Raji cells upon inhibition of translation by cycloheximide; further data indicated that this effect was not correlated with the inhibition of the synthesis of trans-acting protein(s) required for mRNA stability. In the present work, we developed an in vitro mRNA decay assay system to measure HLA-DRA mRNA stability and used inhibitors of protein synthesis affecting different steps of the process of translation in order to discriminate among possible mechanisms determining controlled MHC class II mRNA hydrolysis. We found that HLA-DRA mRNA associated with polysomes derived from cells treated with either puromycin (which causes dispersion of polysomes and accumulation of monosomes) or cycloheximide (which slows down translation causing ribosome stalling) is more rapidly degraded than in the absence of protein synthesis inhibitors. On the basis of our findings, we suggest that arrest of protein synthesis per se exposes the HLA-DRA mRNA molecules to degradative activities co-sedimenting with the polysomal fraction.