Characterization of a splicing mutation in group A xeroderma pigmentosum

Journal Article · Sat Dec 01 00:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.24.9908· OSTI ID:5044264

Satokata, Ichiro; Tanaka, Kiyoji; Miura, Naoyuki; Miyamoto, Iwai; Okada, Yoshio ^[1]; Satoh, Yoshiaki ^[2]; Kondo, Seiji ^[3]

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1. Osaka Univ. (Japan)
2. Tokyo Women's Medical College (Japan) Tokyo Medical and Dental Univ. (Japan)
3. Tokyo Medical and Dental Univ. (Japan)

The molecular basis of group A xeroderma pigmentosum (WP) was investigated by comparison of the nucleotide sequences of multiple clones of the XP group A complementing gene (XPAC) from a patient with group A XP with that of a normal gene. The clones showed a G {r arrow} C substitution at the 3{prime} splice acceptor site of intron 3, which altered the obligatory AG acceptor dinucleotide to AC. Nucleotide sequencing of cDNAs amplified by the polymerase chain reaction revealed that this single base substitution abolishes the canonical 3{prime} splice site, thus creating two abnormally spliced mRNA forms. The larger form is identical with normal mRNA except for a dinucleotide deletion at the 5{prime} end of exon 4. This deletion results in a frameshift with premature translation termination in exon 4. The smaller form has a deletion of the entire exon 3 and the dinucleotide at the 5{prime} end of exon 4. The result of a transfection study provided additional evidence that this single base substitution is the disease-causing mutation. This single base substitution creates a new cleavage site for the restriction nuclease AlwNI. Analysis of AlwNI restriction fragment length polymorphism showed a high frequency of this mutation in Japanese patients with group A XP: 16 of 21 unrelated Japanese patients were homozygous and 4 were heterozygous for this mutation. However, 11 Caucasians and 2 Blacks with group A XP did not have this mutant allele. The polymorphic AlwNI restriction fragments are concluded to be useful for diagnosis of group A XP in Japanese subjects, including prenatal cases and carriers.

• OSTI ID: 5044264
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 87:24; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GENE MUTATIONS
DNA SEQUENCING
XERODERMA PIGMENTOSUM
MOLECULAR BIOLOGY
DNA HYBRIDIZATION
NUCLEASES
PATIENTS
PHOSPHORUS 32
RECOMBINANT DNA
RFLPS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
DISEASES
DNA
ENZYMES
ESTERASES
HYBRIDIZATION
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MUTATIONS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PHOSPHORUS ISOTOPES
RADIOISOTOPES
SKIN DISEASES
STRUCTURAL CHEMICAL ANALYSIS
550201* - Biochemistry- Tracer Techniques