Molecular characterization of severe hemophilia A suggests that about half the mutations are not within the coding regions and splice junctions of the factor VIII gene
| Autor: | Haig H. Kazazian, Laura Kasch, Miyoko Higuchi, Stylianos E. Antonarakis, John A. Phillips, Robert L. Janco, Matthew J. McGinniss, Tina C. Warren, Carol K. Kasper |
|---|---|
| Předmět: |
RNA Splicing
Molecular Sequence Data Gene mutation Biology Hemophilia A medicine.disease_cause Polymerase Chain Reaction Exon Gene mapping hemic and lymphatic diseases Gene duplication Leukocytes medicine Humans Coding region Amino Acid Sequence Codon Gene Genetics Mutation Factor VIII Polymorphism Genetic Multidisciplinary Base Sequence Computers Chromosome Mapping DNA Exons Molecular biology Genes Haplotypes Coagulation Oligonucleotide Probes Research Article |
| Zdroj: | Scopus-Elsevier |
| Popis: | Hemophilia A is an X chromosome-linked disorder resulting from deficiency of factor VIII, an important protein in blood coagulation. A large number of disease-producing mutations have been reported in the factor VIII gene. However, a comprehensive analysis of the mutations has been difficult because of the large gene size, its many scattered exons, and the high frequency of de novo mutations. Recently, we have shown that nearly all mutations resulting in mild-to-moderate hemophilia A can be detected by PCR and denaturing gradient gel electrophoresis (DGGE). In this study, we attempted to discover the mutations causing severe hemophilia A by analyzing 47 unselected patients, 30 of whom had severe hemophilia and 17 of whom had mild-to-moderate disease. Using DGGE as a screening method, we analyzed 99% of the coding region, 94% of the splice junctions, the promoter region, and the polyadenylylation site of the gene. We found the mutation in 16 of 17 (94%) patients with mild-to-moderate disease but in only 16 of 30 (53%) patients with severe hemophilia A. Since DGGE after computer analysis appears to detect all mutations in a given fragment, the lower-than-expected yield of mutations in patients with severe disease is likely not due to failure of the detection method; it is probably due to the presence of mutations in DNA sequences outside the regions studied. Such sequences may include locus-controlling regions, other sequences within introns or outside the gene that are important for its expression, or another gene involved in factor VIII expression that is very closely linked to the factor VIII gene. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|