Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat

P. G. Reddy; M. R. Nemali; M. K. Reddy; M. N. Reddy; P. M. Yuan; S. Yuen; T. G. Laffler; T. Shiroza; H. K. Kuramitsu; N. Usuda; R. L. Chisholm; M. S. Rao; J. K. Reddy

doi:10.1073/pnas.85.23.9081

Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat

P. G. Reddy, M. R. Nemali, M. K. Reddy, M. N. Reddy, P. M. Yuan, S. Yuen, T. G. Laffler, T. Shiroza, H. K. Kuramitsu, N. Usuda, R. L. Chisholm, M. S. Rao, J. K. Reddy

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Abstract

Urate oxidase (UOxase; ureate:oxygen oxidoreductase, EC 1.7.3.3), which catalyzes the oxidation of uric acid to allantoin, is present in most mammals but is absent in humans and certain primates. A cDNA clone for UOxase containing an insert of 1.3 kilobases (kb) was isolated from a λgt11 cDNA library prepared from rat liver mRNA. This recombinant clone with a 1283-nucleotide insert has sequence for 97% of the coding region together with 401 nucleotides of the 3'-untranslated region of the mRNA. The identity of UOxase cDNA clone was verified by analyzing the fusion protein, immunocytochemical localization with epitope-selected antibody, and hybrid-select translation analysis and by comparing sequences of four CNBr-cleaved peptides of the protein. Blot analysis revealed that the probe hybridizes to a single 1.5-kb mRNA species in the rat liver and a transplantable hepatocellular carcinoma. No UOxase mRNA was detected in 11 nonhepatic tissues of rat, suggesting tissue specificity of expression of this UOxase gene. Blot analysis of RNA from livers of rats treated with a peroxisome proliferator showed 2- to 3-fold increase in UOxase mRNA content, whereas the fatty acyl-CoA oxidase mRNA increased over 30-fold. Southern blot analysis of restriction enzyme digests of rat DNA suggests that there is a single copy of UOxase gene. Analysis of human genomic DNA revealed restriction fragments that are homologous to rat UOxase cDNA, although no UOxase mRNA was detected in human liver.

Original language	English (US)
Pages (from-to)	9081-9085
Number of pages	5
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	85
Issue number	23
DOIs	https://doi.org/10.1073/pnas.85.23.9081
State	Published - 1988

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Reddy, P. G., Nemali, M. R., Reddy, M. K., Reddy, M. N., Yuan, P. M., Yuen, S., Laffler, T. G., Shiroza, T., Kuramitsu, H. K., Usuda, N., Chisholm, R. L., Rao, M. S., & Reddy, J. K. (1988). Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat. Proceedings of the National Academy of Sciences of the United States of America, 85(23), 9081-9085. https://doi.org/10.1073/pnas.85.23.9081

@article{3ae1389fb7784a97897fc4446c6d7085,

title = "Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat",

abstract = "Urate oxidase (UOxase; ureate:oxygen oxidoreductase, EC 1.7.3.3), which catalyzes the oxidation of uric acid to allantoin, is present in most mammals but is absent in humans and certain primates. A cDNA clone for UOxase containing an insert of 1.3 kilobases (kb) was isolated from a λgt11 cDNA library prepared from rat liver mRNA. This recombinant clone with a 1283-nucleotide insert has sequence for 97% of the coding region together with 401 nucleotides of the 3'-untranslated region of the mRNA. The identity of UOxase cDNA clone was verified by analyzing the fusion protein, immunocytochemical localization with epitope-selected antibody, and hybrid-select translation analysis and by comparing sequences of four CNBr-cleaved peptides of the protein. Blot analysis revealed that the probe hybridizes to a single 1.5-kb mRNA species in the rat liver and a transplantable hepatocellular carcinoma. No UOxase mRNA was detected in 11 nonhepatic tissues of rat, suggesting tissue specificity of expression of this UOxase gene. Blot analysis of RNA from livers of rats treated with a peroxisome proliferator showed 2- to 3-fold increase in UOxase mRNA content, whereas the fatty acyl-CoA oxidase mRNA increased over 30-fold. Southern blot analysis of restriction enzyme digests of rat DNA suggests that there is a single copy of UOxase gene. Analysis of human genomic DNA revealed restriction fragments that are homologous to rat UOxase cDNA, although no UOxase mRNA was detected in human liver.",

author = "Reddy, {P. G.} and Nemali, {M. R.} and Reddy, {M. K.} and Reddy, {M. N.} and Yuan, {P. M.} and S. Yuen and Laffler, {T. G.} and T. Shiroza and Kuramitsu, {H. K.} and N. Usuda and Chisholm, {R. L.} and Rao, {M. S.} and Reddy, {J. K.}",

year = "1988",

doi = "10.1073/pnas.85.23.9081",

language = "English (US)",

volume = "85",

pages = "9081--9085",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "23",

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Reddy, PG, Nemali, MR, Reddy, MK, Reddy, MN, Yuan, PM, Yuen, S, Laffler, TG, Shiroza, T, Kuramitsu, HK, Usuda, N, Chisholm, RL , Rao, MS & Reddy, JK 1988, 'Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat', Proceedings of the National Academy of Sciences of the United States of America, vol. 85, no. 23, pp. 9081-9085. https://doi.org/10.1073/pnas.85.23.9081

TY - JOUR

T1 - Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase

T2 - Liver-specific expression in the rat

AU - Reddy, P. G.

AU - Nemali, M. R.

AU - Reddy, M. K.

AU - Reddy, M. N.

AU - Yuan, P. M.

AU - Yuen, S.

AU - Laffler, T. G.

AU - Shiroza, T.

AU - Kuramitsu, H. K.

AU - Usuda, N.

AU - Chisholm, R. L.

AU - Rao, M. S.

AU - Reddy, J. K.

PY - 1988

Y1 - 1988

N2 - Urate oxidase (UOxase; ureate:oxygen oxidoreductase, EC 1.7.3.3), which catalyzes the oxidation of uric acid to allantoin, is present in most mammals but is absent in humans and certain primates. A cDNA clone for UOxase containing an insert of 1.3 kilobases (kb) was isolated from a λgt11 cDNA library prepared from rat liver mRNA. This recombinant clone with a 1283-nucleotide insert has sequence for 97% of the coding region together with 401 nucleotides of the 3'-untranslated region of the mRNA. The identity of UOxase cDNA clone was verified by analyzing the fusion protein, immunocytochemical localization with epitope-selected antibody, and hybrid-select translation analysis and by comparing sequences of four CNBr-cleaved peptides of the protein. Blot analysis revealed that the probe hybridizes to a single 1.5-kb mRNA species in the rat liver and a transplantable hepatocellular carcinoma. No UOxase mRNA was detected in 11 nonhepatic tissues of rat, suggesting tissue specificity of expression of this UOxase gene. Blot analysis of RNA from livers of rats treated with a peroxisome proliferator showed 2- to 3-fold increase in UOxase mRNA content, whereas the fatty acyl-CoA oxidase mRNA increased over 30-fold. Southern blot analysis of restriction enzyme digests of rat DNA suggests that there is a single copy of UOxase gene. Analysis of human genomic DNA revealed restriction fragments that are homologous to rat UOxase cDNA, although no UOxase mRNA was detected in human liver.

AB - Urate oxidase (UOxase; ureate:oxygen oxidoreductase, EC 1.7.3.3), which catalyzes the oxidation of uric acid to allantoin, is present in most mammals but is absent in humans and certain primates. A cDNA clone for UOxase containing an insert of 1.3 kilobases (kb) was isolated from a λgt11 cDNA library prepared from rat liver mRNA. This recombinant clone with a 1283-nucleotide insert has sequence for 97% of the coding region together with 401 nucleotides of the 3'-untranslated region of the mRNA. The identity of UOxase cDNA clone was verified by analyzing the fusion protein, immunocytochemical localization with epitope-selected antibody, and hybrid-select translation analysis and by comparing sequences of four CNBr-cleaved peptides of the protein. Blot analysis revealed that the probe hybridizes to a single 1.5-kb mRNA species in the rat liver and a transplantable hepatocellular carcinoma. No UOxase mRNA was detected in 11 nonhepatic tissues of rat, suggesting tissue specificity of expression of this UOxase gene. Blot analysis of RNA from livers of rats treated with a peroxisome proliferator showed 2- to 3-fold increase in UOxase mRNA content, whereas the fatty acyl-CoA oxidase mRNA increased over 30-fold. Southern blot analysis of restriction enzyme digests of rat DNA suggests that there is a single copy of UOxase gene. Analysis of human genomic DNA revealed restriction fragments that are homologous to rat UOxase cDNA, although no UOxase mRNA was detected in human liver.

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

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Isolation and sequence determination of a cDNA clone for rat peroxisomal urate oxidase: Liver-specific expression in the rat

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