CLONING OF THE CANINE BETA-GLUCURONIDASE CDNA, MUTATION IDENTIFICATION IN CANINE MPS VII, AND RETROVIRAL VECTOR-MEDIATED CORRECTION OF MPS VII CELLS

Mucopolysaccharidosis type VII (MPS VII) is an inherit;ed disease resu lting from deficient activity of the lysosomal acid hydrolase beta-glu curonidase (GUSB) and has been reported in humans, mice, cats, and dog s. To characterize canine MPS VII, we have isolated and sequenced the canine GUSB cDNA from normal and affected animals. A single nucleotide substitution was detected in the GUSB cDNA derived from MPS VII dogs, This guanosine to adenine base change at nucleotide position 559 in t he canine cDNA sequence causes an arginine to histidine substitution a t amino acid position 166. introduction of the G to A substitution at position 559 in a mammalian expression vector containing the normal ca nine GUSB cDNA nearly eliminated the GUSB enzymatic activity, demonstr ating that this mutation is the cause of canine MPS VII, A retroviral vector expressing the full-length canine beta-glucuronidase cDNA corre cted the deficiency in MPS VII cells. (C) 1998 Academic Press.