Leukocyte adhesion deficiency. Aberrant splicing of a conserved integrin sequence causes a moderate deficiency phenotype

• Published in: The Journal of biological chemistry 1989-02, Vol.264 (6), p.3588-3595
• Main Authors: Kishimoto, T K, O'Conner, K, Springer, T A
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; DNA - genetics; DNA Probes; DNA-Directed DNA Polymerase - metabolism; Endonucleases; Exons; Fundamental and applied biological sciences. Psychology; Humans; Integrins; Introns; Macromolecular Substances; Male; Membrane Glycoproteins - deficiency; Membrane Glycoproteins - genetics; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Mutation; Phenotype; Protein Precursors - genetics; RNA - genetics; RNA Splicing; RNA, Messenger - genetics; Single-Strand Specific DNA and RNA Endonucleases; Transcription. Transcription factor. Splicing. Rna processing
• ISSN: 0021-9258; 1083-351X

Leukocyte adhesion deficiency (LAD) is a heritable deficiency of the LFA-1, Mac-1, p150,95 family of leukocyte alpha beta heterodimers (the leukocyte integrins). We have studied the defect in patients who synthesize an aberrantly small form of the beta subunit common to all three proteins. S1 nuclease protection showed the presence of a 90-nucleotide mismatch in RNA from patients and relatives, correlating with inheritance of the disease. Use of the Taq polymerase chain reaction to amplify this region of RNA after first strand cDNA synthesis and sequencing showed an in-frame deletion of 90 nucleotides in the extracellular domain. Thus, this highly conserved region, 63% and 53% identical in amino acid sequence to two other beta subunits of the integrin family, is required for association of the beta subunit with alpha subunits. The 90-nucleotide region corresponds to a single exon present in both the normal and patient genome. The patient DNA has a single G to C substitution in the 5' splice site. This results in the direct joining of nonconsecutive exons in an unusual type of abnormal RNA splicing. A small amount of normally spliced message, detected by S1 nuclease protection and Taq polymerase chain reaction, encodes a normal sized beta subunit which is surface-expressed and accounts for the low levels of leukocyte integrin expression observed in these patients, and hence the moderate phenotype.