We have characterized the molecular basis of beta-hexosaminidase A (HEX A) 
deficiency in a patient ascertained through an ophthalmologic examination that 
revealed cherry red spots on his retina. The absence of neurological deficit in 
this child until 3 3/4 years of age indicated residual HEX A must be present. 
Three HEXA mutations, 10T > C (S4P) and 972T > A (V324V) on the maternal allele, 
and 1A > T (M1L) on the paternal allele were identified. The effects of the 
amino acid substitutions on HEX A expressed in COS-7 cells were analyzed; as 
expected, no HEX A activity was associated with the M1L mutation but 
surprisingly, the S4P mutation resulted in 59% of the HEX A activity expressed 
by the wild type cDNA. The effect of the S4P change was much less than that of 
another HEXA mutation, G269S, associated with an adult onset form of G(M2) 
gangliosidosis. This indicated that the S4P change was not the cause of disease 
and suggested that one of the mutations on the maternal allele, 10T > C or 972T 
> A, had its effect at the mRNA level. This was confirmed by Northern blot 
analysis that showed only 7% of the normal level of HEXA mRNA in proband 
fibroblasts. Analysis of the residual mRNA by RT/PCR and sequencing revealed 
normal transcripts from both the maternal and paternal allele, as well as a low 
abundance aberrant transcript from the maternal allele. Sequencing of this 
aberrant transcript revealed a new exon 8 donor site created by the 972T > A 
mutation that resulted in a 17 bp deletion and destabilization of the resulting 
abnormal transcript. The remaining normal mRNA produced from the 972T > A allele 
must account for the delayed onset of clinical symptoms in this child.