A 3'UTR mutation affects beta-globin expression without altering the stability of its fully processed mRNA.

Abstract

Determinants of mRNA stability are frequently positioned in the 3'UTR where they are not subject to disruption by actively translating ribosomes. Two related individuals with beta thalassaemia who carry a beta-globin gene containing a 13 nt deletion in its 3'UTR have recently been described. Its position within the 3'UTR, as well as its relative distance from other known functionally important elements, suggested that the deletion might overlay previously unrecognized determinants of beta-globin mRNA stability. We studied the impact of the Delta13 mutation on beta-globin gene expression in vitro and in vivo. The adverse effect of the Delta13 mutation on beta-globin expression was confirmed in studies utilizing reticulocytes from a betaDelta13 heterozygote, which indicated a sixfold reduction in the relative level of the mutant mRNA. Additional in vitro analysis indicated that the deletion did not affect the capacity of the betaDelta13 mRNA to assemble an mRNA-stabilizing mRNP 'beta-complex'. Unexpectedly, functional tests in both primary erythroid cells and in a transgenic mouse model demonstrated that the betaDelta13 mRNA was fully stable, suggesting that the Delta13 mutation affects accumulation of the fully processed mRNA at an earlier step. Consistent with this, there was a relative excess of unprocessed betaDelta13 mRNA in erythroid progenitors from a betaDelta13 heterozygote. Taken together, these results define a new thalassaemic determinant, which acts to decrease beta-globin mRNA levels by inhibiting the efficiency of nuclear processing events, and suggest a previously unanticipated complexity to the role of the 3'UTR elements in the regulation of beta-globin gene expression.