Isolation, characterization, and expression of cDNA clones that encode rat UDP-galactose: ceramide galactosyltransferase.

Abstract

UDP-galactose:ceramide galactosyltransferase (CGT) (EC. 2.4.1.62) catalyzes the final step in the synthesis of galactocerebroside (GalC), a glycosphingolipid found in high amounts in the myelin sheath. Here, the isolation of rat CGT specific cDNA clones is reported. The CGT sequence contains an open reading frame of 1,623 bp which predicts a protein of M(r) 61,126 Da. In transfection experiments the cDNA was found to confer CGT activity to Chinese hamster ovary cells. In rat brain the developmental expression pattern of CGT mRNA was similar to the myelination profile, whereas the sciatic nerve contained high amounts of CGT message over a long developmental period. CGT mRNA expression in the sciatic nerve was found to drop substantially following nerve injury and recover slowly when compared to the expression of mRNAs specific for the predominant myelin-specific proteins. The absolute amounts of CGT message in sciatic nerve and brain were found to be comparable to those that encode the structural proteins of myelin. Except for low amounts in the kidney, the CGT mRNA was not detected in other tissues examined. Southern blot analysis revealed that the CGT protein is likely encoded by a single, relatively large gene.