HIGH-RESOLUTION MAPPING OF THE GAMMA-AMINOBUTYRIC-ACID RECEPTOR SUBUNIT-BETA-3 AND ALPHA-5 GENE-CLUSTER ON CHROMOSOME-15Q11-Q13, AND LOCALIZATION OF BREAKPOINTS IN 2 ANGELMAN SYNDROME PATIENTS

The gamma-aminobutyric acid (GABA(A)) receptors are a family of ligand-gated chloride channels constituting the major inhibitory neurotransmitter receptors in the nervous system. In order to determine the genomic organization of the GABA(A) receptor beta3 subunit gene (GABRB3) and alpha5 subunit gene (GABRA5) in chromosome 15q11-q13, we have constructed a high-resolution physical map using the combined techniques of field-inversion gel electrophoresis and phage genomic library screening. This map, which covers nearly 1.0 Mb, shows that GABRB3 and GABRA5 are separated by less than 100 kb and are arranged in a head-to-head configuration. GABRB3 encompasses approximately 250 kb, while GABRA5 is contained within 70 kb. This difference in size is due in large part to an intron of 150 kb within GABRB3. We have also identified seven putative CpG islands within a 600-kb interval. Chromosomal rearrangement breakpoints-in one Angelman syndrome (AS) patient with an unbalanced translocation and in another patient with a submicroscopic deletion-are located within the large GABRB3 intron. These findings will facilitate chromosomal walking strategies for cloning the regions disrupted by the DNA rearrangements in these AS patients and will be valuable for mapping new genes to the AS chromosomal region.