FRAMESHIFT AND NONSENSE MUTATIONS IN A HUMAN GENOMIC SEQUENCE HOMOLOGOUS TO A MURINE UDP-GAL-BETA-D-GAL(1,4)-D-GLCNAC ALPHA-(1,3)-GALACTOSYLTRANSFERASE CDNA

Abstract

We have previously isolated a murine UDP-Gal:.beta.-D-Gal(1,4)-D-GlcNac .alpha.(1,3)-galactosyltransferase (.alpha.(1,3)-GT) cDNA (Larsen, R. D., Rajan, V. P., Ruff, M. M., Kukowska-Latallo, J., Cummings, R. D., and Lowe, J.B. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 8227-8231). This enzyme constructs the terminal .alpha.(1,3)-galactosyl linkage within the epitope Gal.alpha.1.fwdarw.3Gal. This epitope is expressed by New World monkeys and many nonprimate mammals but generally not by Old World primates, anthropoid apes, or man. To investigate the molecular bases for the apparent species-specific absence of this enzyme and its oligosaccharide product, we have sequenced a human genomic DNA fragment homologous to the murine .alpha.(1,3)-GT cDNA. This fragment contains a 703-nucleotide region that shares 82% identity with a region of the murine cDNA encoding part of the enzyme''s catalytic domain. The human sequence, however, has suffered deletion of single nucleotides at two separate positions, relative to the murine sequence. These frameshift mutations disrupt the translational reading frame that would otherwise maintain a 76% amino acid sequence identity between the human sequence and the murine .alpha.(1,3)-GT. Moreover, nonsense mutations exist within this disrupted reading frame that would truncate the human polypeptide, relative to the murine enzyme. We therefore propose that this human sequence represents a pseudogene and cannot determine expression of Gal.alpha.1.fwdarw.3Gal epitopes on human cells.