Cloning and chromosomal mapping of the mouse Mgat3 gene encoding N-acetylglucosaminyltransferase III.
Complex and hybrid N-linked carbohydrates synthesized by mammalian cells may possess a N-acetylglucosamine residue known as the bisecting GlcNAc. The transfer of this residue is catalyzed by the enzyme UDP-N-acetylglucosamine:beta-D-mannoside beta 1,4-N-acetylglucosaminyltransferase III (GlcNAc-TIII; EC 2.4.1.144). To begin to investigate biological functions for carbohydrates with a bisected GlcNAc residue, we have cloned and partially characterized the mouse gene (Mgat3) encoding GlcNAc-TIII. A rat GlcNAc-TIII-encoding cDNA was used to isolate clones from a mouse strain 129 Sv liver genomic DNA library. An NsiI genomic DNA fragment containing an ORF with 96% identity to rat GlcNAc-TIII was subcloned into a mammalian expression vector and transfected into Chinese hamster ovary (CHO) cells. The transfectants expressed GlcNAc-TIII activity only when the ORF was in the sense orientation. Southern analysis showed that Mgat3 is present in a single copy in the mouse genome. Mapping by restriction-fragment length polymorphism analysis of backcross progeny located Mgat3 to mouse chromosome 15, at a position homologous with region 22q12.3-q13.1 in the human genome. Northern analyses of adult tissues showed that Mgat3 is expressed at high levels in kidney and brain, and at lower levels in many other tissues.
Bhaumik
M
M
Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA.
Seldin
M F
MF
Stanley
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002874:Chromosome Mapping; D002892:Chromosomes, Human, Pair 22; D003001:Cloning, Molecular; D006224:Cricetinae; D015698:Genomic Library; D006801:Humans; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis* / Q000235:genetics*; D016366:Open Reading Frames; D009928:Organ Specificity; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D011994:Recombinant Proteins; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase