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Molecular basis of multiple UDP-glucuronosyltransferase isoenzyme deficiencies in the hyperbilirubinemic rat (Gunn rat).

Open AccessPublished:December 15, 1991DOI:https://doi.org/10.1016/S0021-9258(18)54390-8
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      The Gunn rat is a mutant strain of Wistar rat which has unconjugated hyperbilirubinemia as a result of the absence of hepatic UDP-glucuronosyltransferase (UDPGT) activity toward bilirubin. The Gunn rat is also deficient in UDPGT activities toward phenol substrates, and also toward digitoxigenin-monodigitoxiside. We have demonstrated that the defect of the isoenzyme for 4-nitrophenol (4NP) in Gunn rat liver arises from a -1 frameshift mutation that removes 115 amino acids from the COOH terminus (Iyanagi, T., Watanabe, T., and Uchiyama, Y. (1989) J. Biol. Chem. 264, 21302-21307). To investigate the molecular basis of defects in other UDPGT isoenzymes, we isolated and sequenced cDNAs from a Gunn rat liver library using mutant 4NP-UDPGT cDNA as a probe. Three novel cDNAs were identified that had identical 3'-regions of 1362 base pairs containing a single-base deletion in the same position as that of the mutant 4NP-UDPGT cDNA. However, their 5'-regions, encoding the substrate-binding domain, showed no more than 40% homology to that of 4NP-UDPGT. These data provide evidence that defects in some UDPGT isoenzymes in the Gunn rat are caused by a single mutation that results in the formation of a common truncated COOH terminus. Furthermore, the data also suggest that these mRNAs are transcribed from a single gene and that the 5'-exons are transcribed independently and differentially spliced to common 3'-exons encoding the conserved domain.

      REFERENCES

        • Gunn C.H.
        J. Heredity. 1938; 29: 137-139
        • Lathe G.H.
        • Walker M.
        Biochem. J. 1957; 67: 9P
        • Cornelius E.E.
        • Arias I.M.
        Am. J. Pathol. 1972; 69: 369-372
        • Watkins J.B.
        • Klaassen C.D.
        Drug. Metab. Dispos. 1982; 10: 590-594
        • Mackenzie P.I.
        • Owens I.S.
        Biochem. Pharmacol. 1983; 32: 3777-3781
        • Drucker W.D.
        Proc. Soc. Exp. Biol. Med. 1968; 129: 308-312
        • Jackson M.R.
        • Burchell B.
        Nucleic Acids Res. 1986; 14: 779-795
        • Mackenzie P.I.
        J. Biol. Chem. 1986; 261: 6119-6125
        • Mackenzie P.I.
        J. Biol. Chem. 1986; 261: 14112-14117
        • Mackenzie P.I.
        J. Biol. Chem. 1987; 262: 9744-9749
        • Iyanagi T.
        • Haniu M.
        • Sogawa K.
        • Fujii-Kuriyama Y.
        • Watanabe S.
        • Shively J.E.
        • Anan K.F.
        J. Biol. Chem. 1986; 261: 15607-15614
        • Harding D.
        • Fournel-Gigleux S.
        • Jackson M.R.
        • Burchell B.
        Proc. Natl. Acad. Sci. U. S. A. 1988; 85: 8381-8385
        • Iyanagi T.
        • Watanabe T.
        • Uchiyama Y.
        J. Biol. Chem. 1989; 264: 21302-21307
        • Nagai F.
        • Takahashi M.
        • Homma H.
        • Tanase H.
        • Matsui M.
        J. Pharmacobio-Dyn. 1987; 10: 421-426
        • Chirgwin J.M.
        • Przybyla A.E.
        • MacDnald R.J.
        • Rutter W.J.
        Biochemistry. 1979; 18: 5294-5299
        • Huynh T.V.
        • Young R.A.
        • Davis R.W.
        Glover D.M. DNA Cloning I. IRL Press Ltd., Oxford, Great Britain1985: 49-78
        • Sanger F.
        • Nickleu S.
        • Goulson A.R.
        Proc. Natl. Acad. Sci. U. S. A. 1977; 74: 5463-5467
        • McMaster G.K.
        • Carmichael G.G.
        Proc. Natl. Acad. Sci. U. S. A. 1977; 74: 4835-4838
        • Sambook J.
        • Fritsch E.F.
        • Maniatis T.
        Molecular Cloning: A Laboratory Mannual. 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY1989
        • Kozak M.
        Nucleic Acids Res. 1984; 12: 857-872
        • Sato H.
        • Koiwai O.
        • Tanabe K.
        • Kashiwamata S.
        Biochem. Biophys. Res. Commun. 1990; 169: 260-264
        • Burchell B.
        • Nebert D.W.
        • Nelson D.R.
        • Bock K.W.
        • Iyanagi T.
        • Jansen P.L.M.
        • Lancet D.
        • Mulder G.J.
        • Roy Chowdhury J.
        • Siest G.
        • Tephly T.R.
        • Mackenzie P.I.
        DNA Cell Biol. 1991; (in press)
        • Coffman B.L.
        • Tephly T.R.
        • Irshaid Y.M.
        • Green M.D.
        • Smith C.
        • Jackson M.R.
        • Wooster R.
        • Burchell B.
        Arch. Biochem. Biophys. 1990; 281: 170-175
        • Iyanagi T.
        Siest G. Magdalou J. Burchell B. Cellular and Molecular Aspects of Glucuronidation. 173. John Libbey & Co. Ltd., London1988: 3-11
        • Falany C.N.
        • Green M.D.
        • Swain E.
        • Tephly T.R.
        Biochem. J. 1986; 238: 65-73
        • ElAwaday M.
        • Roy Chowdhury J.
        • Kesari K.
        • van Es H.H.
        • Jansen P.L.M.
        • Lederstein M.
        • Arias I.M.
        • Roy Chowdhury N.
        J. Biol. Chem. 1990; 265: 10752-10758
        • Mount S.M.
        Nucleic Acids Res. 1982; 10: 459-472
        • Mackenzie P.I.
        • Rodbourn L.
        J. Biol. Chem. 1990; 265: 11328-11332
        • Mackenzie P.I.
        J. Biol. Chem. 1990; 265: 3432-3435
        • Harding D.
        • Jackson M.R.
        • Wooster R.
        • Fournel-Gigleux S.
        • Burchell B.
        Siest G. Magdalou J. Burchell B. Cellular and Molecular Aspects of Glucuronidation. 173. John Libbey & Co. Ltd., London1988: 13-20
        • Ritter J.K.
        • Crawford J.M.
        • Owens I.S.
        J. Biol. Chem. 1991; 266: 1043-1047
        • van Es H.H.
        • Goldhoorn B.G.
        • Paul-Abrahamse M.
        • Elferink P.J.O.
        • Jansen P.L.M.
        J. Clin. Invest. 1990; 85: 1199-1205