A family of human beta4-galactosyltransferases. Cloning and expression of two novel UDP-galactose:beta-n-acetylglucosamine beta1, 4-galactosyltransferases, beta4Gal-T2 and beta4Gal-T3.
BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of the human UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferase, designated beta4Gal-T1, revealed a large number of ESTs with identical as well as similar sequences. ESTs with sequences similar to that of beta4Gal-T1 could be grouped into at least two non-identical sequence sets. Analysis of the predicted amino acid sequence of the novel ESTs with beta4Gal-T1 revealed conservation of short sequence motifs as well as cysteine residues previously shown to be important for the function of beta4Gal-T1. The likelihood that the identified ESTs represented novel galactosyltransferase genes was tested by cloning and sequencing of the full coding region of two distinct genes, followed by expression. Expression of soluble secreted constructs in the baculovirus system showed that these genes represented genuine UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferases, thus designated beta4Gal-T2 and beta4Gal-T3. Genomic cloning of the genes revealed that they have identical genomic organizations compared with beta4Gal-T1. The two novel genes were located on 1p32-33 and 1q23. The results demonstrate the existence of a family of homologous galactosyltransferases with related functions. The existence of multiple beta4-galactosyltransferases with the same or overlapping functions may be relevant for interpretation of biological functions previously assigned to beta4Gal-T1.
Almeida
R
R
School of Dentistry, University of Copenhagen, Norre Allé 20, DK-2200 Copenhagen N, Denmark.
Amado
M
M
David
L
L
Levery
S B
SB
Holmes
E H
EH
Merkx
G
G
van Kessel
A G
AG
Rygaard
E
E
Hassan
H
H
Bennett
E
E
Clausen
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002240:Carbohydrate Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000737:chemistry / Q000235:genetics*; D009844:Oligosaccharides / Q000096:biosynthesis; D011092:Polyethylene Glycols / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D016324:Sequence Tagged Sites
D009844:Oligosaccharides; D011092:Polyethylene Glycols; D009237:N-Acetyllactosamine Synthase