Identification and localization of mutations at the Lesch-Nyhan locus by ribonuclease A cleavage.

TitleIdentification and localization of mutations at the Lesch-Nyhan locus by ribonuclease A cleavage.
Publication TypeJournal Article
Year of Publication1987
AuthorsGibbs, RA, Caskey, CT
JournalScience
Volume236
Issue4799
Pagination303-5
Date Published1987 Apr 17
ISSN0036-8075
KeywordsCell Line, Chromosome Deletion, HeLa Cells, Humans, Hypoxanthine Phosphoribosyltransferase, Lesch-Nyhan Syndrome, Mutation, Nucleic Acid Hybridization, Ribonuclease, Pancreatic, RNA, Messenger
Abstract

Many mutations leading to human disease are the result of single DNA base pair changes that cannot be identified by Southern analysis. This has prompted the development of alternative assays for point mutation detection. The recently described ribonuclease A cleavage procedure, with a polyuridylic acid-paper affinity chromatography step, has been used to identify the mutational lesions in the hypoxanthine phosphoribosyltransferase (HPRT) messenger RNAs of patients with Lesch-Nyhan syndrome. Distinctive ribonuclease A cleavage patterns were identified in messenger RNA from 5 of 14 Lesch-Nyhan patients who were chosen because no HPRT Southern or Northern blotting pattern changes had been found. This approach now allows HPRT mutation detection in 50 percent of the cases of Lesch-Nyhan syndrome. The polyuridylic acid-paper affinity procedure provides a general method for analysis of low abundance messenger RNAs.

Alternate JournalScience
PubMed ID3563511