A 5' Splice-region G----C Mutation in Exon 1 of the Human Beta-globin Gene Inhibits Pre-mRNA Splicing: a Mechanism for Beta+-thalassemia

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1989 Feb 1

PMID 2915972

Citations 17

Authors

M Vidaud

R Gattoni

J Stevenin

D Vidaud

S Amselem

J Chibani

J Rosa

M Goossens

Affiliations

Soon will be listed here.

Abstract

We have characterized a Mediterranean beta-thalassemia allele containing a sequence change at codon 30 that alters both beta-globin pre-mRNA splicing and the structure of the hemoglobin product. Presumably, this G----C transversion at position -1 of intron 1 reduces severely the utilization of the normal 5' splice site since the level of the Arg----Thr mutant hemoglobin (designated hemoglobin Kairouan) found in the erythrocytes of the patient is very low (2% of total hemoglobin). Since no natural mutations of the guanine located at position -1 of the CAG/GTAAGT consensus sequence had been isolated previously, we investigated the role of this nucleotide in the constitution of an active 5' splice site by studying the splicing of the pre-mRNA in cell-free extracts. We demonstrate that correct splicing of the mutant pre-mRNA is 98% inhibited. Our results provide further insights into the mechanisms of pre-mRNA maturation by revealing that the last residue of the exon plays a role at least equivalent to that of the intron residue at position +5.

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