Beta+45 G --> C: a novel silent beta-thalassaemia mutation, the first in the Kozak sequence

A family from the Southeast of Italy was found to have a novel beta-globin mutant, beta+45 G ->C, with the features of a silent beta-thalassaemia mutation. It was asymptomatic in two heterozygotes, but its interaction with the severe thalassaemia mutation beta-IVS-II-654 C->T worsened the haematological and biosynthetic phenotype in two compound heterozygotes; moreover, another compound heterozygote, who was also heterozygote for the aaa anti3.7, suffered from thalassaemia intermedia. The mutation was found associated in cis with the IVS-II-754 T->C substitution, which did not lead to abnormally spliced mRNA. Furthermore, the amount of beta+45 mRNA was the same as the beta A mRNA in the reticulocytes of the carriers. In vitro transcription/translation experiments demonstrated that the beta+45 G->C decreased the efficiency of translation of the beta-globin chain by about 30%: this slight impairment was consistent with the observed clinical phenotype. The beta+45 G->C is the first mutation found in the Kozak sequence (GACACCATGG) of the beta-globin gene and the first one at the position -6 upstream the ATG. The Kozak consensus sequence plays a major role in the initiation of translation process. The present finding supports the hypothesis that the G in position -6 is important in this process.