Piscine UDP-glucuronosyltransferase 1B.
Glucuronidation is an important detoxification pathway for organic pollutants in fish. We report here the isolation and characterisation of UDP-glucuronosyltransferases (UGT) genes from the closely related marine flatfish, plaice (Pleuronectes platessa) and flounder (Platichthys flesus). The deduced amino acid sequences share greater similarity with mammalian UGT1 family genes than UGT2 genes (44-47% and 39-40% amino acid identity, respectively) and have been designated UGT1B. Both plaice and flounder UGT1B mRNAs are expressed in all tissues and are most highly expressed in liver, with high levels in intestine, gill, kidney and adipose tissue and much lower levels in muscle, heart and brain. Plaice UGT1B mRNA is undetectable in gametes or fertilised eggs and there is a large increase in expression between gastrulation and myotome formation after which levels decline some 5-10-fold. Flounder UGT1B mRNA was increased in liver after intraperitoneal injection of Arochlor 1254 or lindane (gamma-hexachlorocyclohexane), but not after perflourooctanoic acid or 3-methylcholanthrene treatment. In isolated flounder hepatocytes UGT1B mRNA was increased after exposure to benzo(a)pyrene but not by 17alpha-ethynylestradiol. Expression of a cDNA for plaice UGT1B in cos7 cells resulted in higher 1-naphthol conjugation in cell homogenates compared to steroid conjugation, whilst bilirubin and bile acid conjugation were undetectable. This indicates that the plaice gene codes for the phenol-conjugating UGT previously purified in our laboratory from this species and that it is likely to play a major role in the detoxification of polyaromatic hydrocarbons in flatfish. Its role in development is unknown. UGT1B genes are also present in pufferfish (Tetraodon nigroviridis) and zebrafish (Danio rerio) genomes, but they differ in their genic organisation. Pufferfish possess multiple (repeated) complete UGT1 genes and Southern blots indicate that the homologous plaice UGT1B gene may also be organised in this way. In contrast, zebrafish appear to have two UGT1 loci whose sequences and intron/exon structures are closely related to that of plaice, however, the organisation of these genes is similar to the mammalian UGT1 family since each has multiple repeated exon 1's which are alternatively spliced to a common set of exons encoding the aglycone binding domain. Taken together with evidence from phylogenetic comparison of fish sequences with UGT1 and UGT2 families in mammals, we suggest these homologous fish UGTs should all be included within the vertebrate UGT1 family and designated as UGT1B.
Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK.
Michael J
MJ
Leaver
Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK.
Joy
J
Wright
Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK.
Paul
P
Hodgson
Fisheries Research Institute, National Agricultural Research Foundation, Nea Peramos, 64007 Kavala, Greece.
Evridiki
E
Boukouvala
Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK. Electronic address: sgg1@stir.ac.uk.
Stephen G
SG
George
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002210:Caprylates / Q000494:pharmacology; D020111:Chlorodiphenyl (54% Chlorine) / Q000494:pharmacology; D005260:Female; D005432:Flounder / Q000235:genetics / Q000378:metabolism*; D005466:Fluorocarbons / Q000494:pharmacology; D014453:Glucuronosyltransferase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D022781:Hepatocytes / Q000201:enzymology; D001556:Hexachlorocyclohexane / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014874:Water Pollutants, Chemical / Q000494:pharmacology
D002210:Caprylates; D005466:Fluorocarbons; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical; D020111:Chlorodiphenyl (54% Chlorine); D008748:Methylcholanthrene; D001556:Hexachlorocyclohexane; C023036:perfluorooctanoic acid; D014453:Glucuronosyltransferase