Cloning, expression, and characterization of a novel UDP-galactose:beta-N-acetylglucosamine beta1,3-galactosyltransferase (beta3Gal-T5) responsible for synthesis of type 1 chain in colorectal and pancreatic epithelia and tumor cells derived therefrom.
The sialyl Lewis a antigen is a well known tumor marker, CA19-9, which is frequently elevated in the serum in gastrointestinal and pancreatic cancers. UDP-galactose:N-acetylglucosamine beta1, 3-galactosyltransferase(s) (beta3Gal-Ts) are required for the synthesis of the sialyl Lewis a epitope. In the present study, a novel beta3Gal-T, named beta3Gal-T5, was isolated from a Colo205 cDNA library using a degenerate primer strategy based on the amino acid sequences of the four human beta3Gal-T genes cloned to date. Transfection experiments demonstrated that HCT-15 cells transfected with the beta3Gal-T5 gene expressed all the type 1 Lewis antigens. In gastrointestinal and pancreatic cancer cell lines, the amounts of beta3Gal-T5 transcripts were quite well correlated with the amounts of the sialyl Lewis a antigens. The beta1,3Gal-T activity toward agalacto-lacto-N-neotetraose was also well correlated with the amounts of beta3Gal-T5 transcripts in a series of cultured cancer cells, and in Namalwa and HCT-15 cells transfected with the beta3Gal-T5 gene. Thus, the beta3Gal-T5 gene is the most probable candidate responsible for the synthesis of the type 1 Lewis antigens in gastrointestinal and pancreatic epithelia and tumor cells derived therefrom. In addition, beta3Gal-T5 is a key enzyme that determines the amounts of the type 1 Lewis antigens including the sialyl Lewis a antigen.
Isshiki
S
S
Division of Cell Biology, Institute of Life Science, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, Japan.
Togayachi
A
A
Kudo
T
T
Nishihara
S
S
Watanabe
M
M
Kubota
T
T
Kitajima
M
M
Shiraishi
N
N
Sasaki
K
K
Andoh
T
T
Narimatsu
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D018395:CA-19-9 Antigen; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D018076:DNA, Complementary; D005732:Gangliosides / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D018395:CA-19-9 Antigen; D018076:DNA, Complementary; D005732:Gangliosides; D012333:RNA, Messenger; C064564:sialyl Le(a) ganglioside; D017855:Transferases (Other Substituted Phosphate Groups); C037363:UDPgalactose - UDP-N-acetylglucosamine galactosephosphotransferase