GT to AT Transition at a Splice Donor Site Causes Skipping of the Preceding Exon in Phenylketonuria

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1987 Jul 24

PMID 3615198

Citations 47

Authors

J Marvit

A G DiLella

K Brayton

F D Ledley

K J Robson

S L Woo

Affiliations

Soon will be listed here.

Abstract

Classical Phenylketonuria (PKU) is an autosomal recessive human genetic disorder caused by a deficiency of hepatic phenylalanine hydroxylase (PAH). We isolated several mutant PAH cDNA clones from a PKU carrier individual and showed that they contained an internal 116 base pair deletion, corresponding precisely to exon 12 of the human chromosomal PAH gene. The deletion causes the synthesis of a truncated protein lacking the C-terminal 52 amino acids. Gene transfer and expression studies using the mutant PAH cDNA indicated that the deletion abolishes PAH activity in the cell as a result of protein instability. To determine the molecular basis of the deletion, the mutant chromosomal PAH gene was isolated from this individual and shown to contain a GT-- greater than AT substitution at the 5' splice donor site of intron 12. Thus, the consequence of the splice donor site mutation in the human liver is the skipping of the preceding exon during RNA splicing.

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References

Friedman P, Kaufman S, Kang E . Nature of the molecular defect in phenylketonuria and hyperphenylalaninaemia. Nature. 1972; 240(5377):157-9. DOI: 10.1038/240157a0. View

DiLella A, Kwok S, Ledley F, Marvit J, Woo S . Molecular structure and polymorphic map of the human phenylalanine hydroxylase gene. Biochemistry. 1986; 25(4):743-9. DOI: 10.1021/bi00352a001. View

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

Maniatis T, Hardison R, Lacy E, Lauer J, OConnell C, Quon D . The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978; 15(2):687-701. DOI: 10.1016/0092-8674(78)90036-3. View

Birnboim H, DOLY J . A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979; 7(6):1513-23. PMC: 342324. DOI: 10.1093/nar/7.6.1513. View

Woo S . A sensitive and rapid method for recombinant phage screening. Methods Enzymol. 1979; 68:389-95. DOI: 10.1016/0076-6879(79)68028-x. View

Maxam A, Gilbert W . Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980; 65(1):499-560. DOI: 10.1016/s0076-6879(80)65059-9. View

Scriver C, Clow C . Phenylketonuria: epitome of human biochemical genetics (first of two parts). N Engl J Med. 1980; 303(23):1336-42. DOI: 10.1056/NEJM198012043032305. View

Scriver C, Clow C . Phenylketonuria and other phenylalanine hydroxylation mutants in man. Annu Rev Genet. 1980; 14:179-202. DOI: 10.1146/annurev.ge.14.120180.001143. View

10.

Chu G, Sharp P . SV40 DNA transfection of cells in suspension: analysis of efficiency of transcription and translation of T-antigen. Gene. 1981; 13(2):197-202. DOI: 10.1016/0378-1119(81)90008-1. View

11.

Grosveld F, Dahl H, de Boer E, Flavell R . Isolation of beta-globin-related genes from a human cosmid library. Gene. 1981; 13(3):227-37. DOI: 10.1016/0378-1119(81)90028-7. View

12.

Breathnach R, Chambon P . Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981; 50:349-83. DOI: 10.1146/annurev.bi.50.070181.002025. View

13.

Guttler F, Ledley F, Lidsky A, DiLella A, Sullivan S, Woo S . Correlation between polymorphic DNA haplotypes at phenylalanine hydroxylase locus and clinical phenotypes of phenylketonuria. J Pediatr. 1987; 110(1):68-71. DOI: 10.1016/s0022-3476(87)80290-1. View

14.

DiLella A, Marvit J, Lidsky A, Guttler F, Woo S . Tight linkage between a splicing mutation and a specific DNA haplotype in phenylketonuria. Nature. 1986; 322(6082):799-803. DOI: 10.1038/322799a0. View

15.

Mitchell P, Urlaub G, Chasin L . Spontaneous splicing mutations at the dihydrofolate reductase locus in Chinese hamster ovary cells. Mol Cell Biol. 1986; 6(6):1926-35. PMC: 367730. DOI: 10.1128/mcb.6.6.1926-1935.1986. View

16.

Woo S, DiLella A, Marvit J, Ledley F . Molecular basis of phenylketonuria and potential somatic gene therapy. Cold Spring Harb Symp Quant Biol. 1986; 51 Pt 1:395-401. DOI: 10.1101/sqb.1986.051.01.047. View

17.

DiLella A, Woo S . Cloning large segments of genomic DNA using cosmid vectors. Methods Enzymol. 1987; 152:199-212. DOI: 10.1016/0076-6879(87)52021-3. View

18.

DiLella A, Woo S . Hybridization of genomic DNA to oligonucleotide probes in the presence of tetramethylammonium chloride. Methods Enzymol. 1987; 152:447-51. DOI: 10.1016/0076-6879(87)52052-3. View

19.

Choo K, Cotton R, Jennings I, Danks D . Observations indicating the nature of the mutation in phenylketonuria. J Inherit Metab Dis. 1980; 2(4):79-84. DOI: 10.1007/BF01805662. View

20.

Mount S . A catalogue of splice junction sequences. Nucleic Acids Res. 1982; 10(2):459-72. PMC: 326150. DOI: 10.1093/nar/10.2.459. View