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Gene Review

ECs0265  -  xanthine-guanine phosphoribosyltransferase

Escherichia coli O157:H7 str. Sakai

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Disease relevance of ECs0265

  • Transfection of cultured monkey kidney cells with recombinant DNA constructed with a cloned Escherichia coli gene that codes for xanthine-guanine phosphoribosyltransferase and several different SV40 DNA-based vectors, results in the synthesis of readily measurable quantities of the bacterial enzyme [1].
  • Northern blot hybridisation analysis of cytoplasmic RNA from pJ-gpt-transformed cells revealed the presence of an EcoRI J DNA complementary RNA species of the same size as the EBV DNA-encoded small RNAs found in EBV-transformed cells [2].
  • Transformation of PCC4 mouse teratocarcinoma stem cells was obtained using a dominant selective marker, the enzyme xanthine-guanine phosphoribosyltransferase (XGPRT), coded by the bacterial Eco.gpt gene placed under the control of the early SV40 genes in the vector pSV2gpt [3].
  • Analyses of the gpt-specific mRNA's produced during infection of CV1 cells indicate that in addition to the mRNA's expected on the basis of known simian virus 40 RNA splicing patterns, there is a novel SV40-gpt hybrid mRNA [4].
  • We have demonstrated the method by isolating a pseudo-wild type revertant virus and a simple deletion mutant virus from a recombinant vaccinia virus with gpt inserted into the vaccinia virus gene encoding the major 35,000-Da secretory protein [5].

High impact information on ECs0265

  • All of them had the ability to convert [14C]xanthine to xanthine monophosphate. pSV2gpt sequences were present and associated with high mol. wt. cellular DNA. pSV2gpt sequences and XGPRT activity were both conserved in the three clones that were propagated in non-selective medium for 30 generations [3].
  • Five independent Eco.gpt-transformed PCC4 cell lines were propagated in selective medium and assayed for XGPRT activity [3].
  • This study utilized four xanthine guanine phosphoribosyltransferase-negative mutant lines to assess the frequency of mutation induced by thymidine at guanine residues in four sequence contexts: the 5' and 3' guanine residues of a GG doublet, the middle guanine residue of a GGG triplet, and the 3' guanine residue of a GGGG quartet [6].
  • This paper describes the structure of the messenger ribonucleic acids (mRNA's) formed during the expression of gpt and an unexpected feature of the nucleotide sequence in the gpt DNA segment [4].
  • This sequence was derived from a region of the Escherichia coli xanthine-guanine phosphoribosyltransferase gene where position 4 is a site frequently mutated by N-methyl-N'-nitrosourea as compared to sites 1-3 [7].

Chemical compound and disease context of ECs0265

  • In contrast to mammalian cells, which do not efficiently use xanthine for purine nucleotide synthesis, cells that produce E. coli XGPRT can synthesize GMP from xanthine via XMP [8].
  • The method depends on the specific inability of a recombinant vaccinia virus expressing the Escherichia coli guanine phosphoriboxyltransferase gene (gpt) to form plaques on a hypoxanthine-guanine phosphoribosyltransferase-negative line of mouse fibroblasts in the presence of 6-thioguanine [5].
  • Passage to nonselective media transiently alters growth of mycophenolic acid-resistant mammalian cells expressing the escherichia coli xanthine-guanine phosphoribosyltransferase gene: implications for sequential selection strategies [9].
  • The Escherichia coli xanthine-guanine phosphoribosyltransferase gene (Ecogpt) rescues mammalian cells from inhibition of purine nucleotide biosynthesis by mycophenolic acid (MPA) [9].
  • We constructed a retrovirus vector, DC/SV6S31GPT, which carries both the Escherichia coli xanthine-guanine phosphoribosyltransferase gene and the mutated Serine 31 DHFR gene [10].

Biological context of ECs0265

  • Plasmid (pLB206) DNA was introduced into human KB cells by Ca2+-mediated DNA transfection, and transformants were selected in medium containing xanthine, aminopterin, and mycophenolic acid, as a consequence of expression of the dominant, selectable XGPRT gene [11].
  • The structure of free XGPRT at 2.25 A resolution confirms the flexibility of residues in and around a mobile loop identified in other PRTases and shows that the cis-peptide conformation of Arg37 at the active site is maintained in the absence of bound ligands [12].
  • The hybrid cells are deficient in hypoxanthine guanine phosphoribosyltransferase (HGPRT) and contain human chromosome 2, marked with Ecogpt, an E. coli gene for xanthine guanine phosphoribosyltransferase [13].
  • With the aim of producing nonviral shuttle vectors for mammalian cells, we have constructed mouse mitochondrial DNA derivatives comprising the xanthine-guanine phosphoribosyltransferase gene as a selectable marker [14].

Associations of ECs0265 with chemical compounds


Analytical, diagnostic and therapeutic context of ECs0265


  1. Expression of a bacterial gene in mammalian cells. Mulligan, R.C., Berg, P. Science (1980) [Pubmed]
  2. Transfer of the Epstein-Barr virus genes coding for small RNAs to human lymphoid cells with a vector carrying a dominant selectable marker. Rymo, L. EMBO J. (1983) [Pubmed]
  3. Stable transformation of mouse teratocarcinoma stem cells with the dominant selective marker Eco.gpt and retention of their developmental potentialities. Bucchini, D., Lasserre, C., Kunst, F., Lovell-Badge, R., Pictet, R., Jami, J. EMBO J. (1983) [Pubmed]
  4. Factors governing the expression of a bacterial gene in mammalian cells. Mulligan, R.C., Berg, P. Mol. Cell. Biol. (1981) [Pubmed]
  5. Reverse guanine phosphoribosyltransferase selection of recombinant vaccinia viruses. Isaacs, S.N., Kotwal, G.J., Moss, B. Virology (1990) [Pubmed]
  6. Thymidine-induced mutations in mammalian cells: sequence specificity and implications for mutagenesis in vivo. Kresnak, M.T., Davidson, R.L. Proc. Natl. Acad. Sci. U.S.A. (1992) [Pubmed]
  7. Formation of O6-methyldeoxyguanosine at specific sites in a synthetic oligonucleotide designed to resemble a known mutagenic hotspot. Richardson, F.C., Boucheron, J.A., Skopek, T.R., Swenberg, J.A. J. Biol. Chem. (1989) [Pubmed]
  8. Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Mulligan, R.C., Berg, P. Proc. Natl. Acad. Sci. U.S.A. (1981) [Pubmed]
  9. Passage to nonselective media transiently alters growth of mycophenolic acid-resistant mammalian cells expressing the escherichia coli xanthine-guanine phosphoribosyltransferase gene: implications for sequential selection strategies. Drews, R.E., Kolker, M.T., Sachar, D.S., Moran, C.P., Schnipper, L.E. Anal. Biochem. (1996) [Pubmed]
  10. Purine salvage rescue by xanthine-guanine phosphoribosyltransferase (XGPRT) potentiates methotrexate resistance conferred by transfer of a mutated dihydrofolate reductase gene. Mineishi, S., Nakahara, S., Takebe, N., Zhao, S.C., Banerjee, D., Bertino, J.R. Cancer Gene Ther. (1998) [Pubmed]
  11. Expression of adenovirus E1a and E1b gene products and the Escherichia coli XGPRT gene in KB cells. Babiss, L.E., Young, C.S., Fisher, P.B., Ginsberg, H.S. J. Virol. (1983) [Pubmed]
  12. Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase. Vos, S., Parry, R.J., Burns, M.R., de Jersey, J., Martin, J.L. J. Mol. Biol. (1998) [Pubmed]
  13. A genetic assay for aneuploidy: quantitation of chromosome loss using a mouse/human monochromosomal hybrid cell line. Sandhu, S.S., Gudi, R., Athwal, R.S. Mutat. Res. (1988) [Pubmed]
  14. Shuttling of integrated vectors from mammalian cells to E. coli is mediated by head-to-tail multimeric inserts. Lutfalla, G., Blanc, H., Bertolotti, R. Somat. Cell Mol. Genet. (1985) [Pubmed]
  15. Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors. Falkner, F.G., Moss, B. J. Virol. (1988) [Pubmed]
  16. Purification and characterization of Escherichia coli xanthine-guanine phosphoribosyltransferase produced by plasmid pSV2gpt. Deo, S.S., Tseng, W.C., Saini, R., Coles, R.S., Athwal, R.S. Biochim. Biophys. Acta (1985) [Pubmed]
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