• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Nov 1, 1995; 14(21): 5209–5215.
PMCID: PMC394630

Targeted disruption of the housekeeping gene encoding glucose 6-phosphate dehydrogenase (G6PD): G6PD is dispensable for pentose synthesis but essential for defense against oxidative stress.


Glucose 6-phosphate dehydrogenase (G6PD) is a housekeeping enzyme encoded in mammals by an X-linked gene. It has important functions in intermediary metabolism because it catalyzes the first step in the pentose phosphate pathway and provides reductive potential in the form of NADPH. In human populations, many mutant G6PD alleles (some present at polymorphic frequencies) cause a partial loss of G6PD activity and a variety of hemolytic anemias, which vary from mild to severe. All these mutants have some residual enzyme activity, and no large deletions in the G6PD gene have ever been found. To test which, if any, function of G6PD is essential, we have disrupted the G6PD gene in male mouse embryonic stem cells by targeted homologous recombination. We have isolated numerous clones, shown to be recombinant by Southern blot analysis, in which G6PD activity is undetectable. We have extensively characterized individual clones and found that they are extremely sensitive to H2O2 and to the sulfydryl group oxidizing agent, diamide. Their markedly impaired cloning efficiency is restored by reducing the oxygen tension. We conclude that G6PD activity is dispensable for pentose synthesis, but is essential to protect cells against even mild oxidative stress.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • D'Urso M, Mareni C, Toniolo D, Piscopo M, Schlessinger D, Luzzatto L. Regulation of glucose 6-phosphate dehydrogenase expression in CHO-human fibroblast somatic cell hybrids. Somatic Cell Genet. 1983 Jul;9(4):429–443. [PubMed]
  • Fraenkel DG. Selection of Escherichia coli mutants lacking glucose-6-phosphate dehydrogenase or gluconate-6-phosphate dehydrogenase. J Bacteriol. 1968 Apr;95(4):1267–1271. [PMC free article] [PubMed]
  • Gorry P, Lufkin T, Dierich A, Rochette-Egly C, Décimo D, Dollé P, Mark M, Durand B, Chambon P. The cellular retinoic acid binding protein I is dispensable. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9032–9036. [PMC free article] [PubMed]
  • Gray GR, Stamatoyannopoulos G, Naiman SC, Kliman MR, Klebanoff SJ, Austin T, Yoshida A, Robinson GC. Neutrophil dysfunction, chronic granulomatous disease, and non-spherocytic haemolytic anaemia caused by complete deficiency of glucose-6-phosphate dehydrogenase. Lancet. 1973 Sep 8;2(7828):530–534. [PubMed]
  • Hooper M, Hardy K, Handyside A, Hunter S, Monk M. HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells. Nature. 1987 Mar 19;326(6110):292–295. [PubMed]
  • Kazazian HH., Jr The thalassemia syndromes: molecular basis and prenatal diagnosis in 1990. Semin Hematol. 1990 Jul;27(3):209–228. [PubMed]
  • Kletzien RF, Harris PK, Foellmi LA. Glucose-6-phosphate dehydrogenase: a "housekeeping" enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J. 1994 Feb;8(2):174–181. [PubMed]
  • Kosower NS, Kosower EM, Wertheim B, Correa WS. Diamide, a new reagent for the intracellular oxidation of glutathione to the disulfide. Biochem Biophys Res Commun. 1969 Nov 6;37(4):593–596. [PubMed]
  • Kuehn MR, Bradley A, Robertson EJ, Evans MJ. A potential animal model for Lesch-Nyhan syndrome through introduction of HPRT mutations into mice. Nature. 1987 Mar 19;326(6110):295–298. [PubMed]
  • Laird PW, Zijderveld A, Linders K, Rudnicki MA, Jaenisch R, Berns A. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 1991 Aug 11;19(15):4293–4293. [PMC free article] [PubMed]
  • Larsen F, Gundersen G, Lopez R, Prydz H. CpG islands as gene markers in the human genome. Genomics. 1992 Aug;13(4):1095–1107. [PubMed]
  • Levy HR. Glucose-6-phosphate dehydrogenases. Adv Enzymol Relat Areas Mol Biol. 1979;48:97–192. [PubMed]
  • Lobo Z, Maitra PK. Pentose phosphate pathway mutants of yeast. Mol Gen Genet. 1982;185(2):367–368. [PubMed]
  • Luzzatto L, Testa U. Human erythrocyte glucose 6-phosphate dehydrogenase: structure and function in normal and mutant subjects. Curr Top Hematol. 1978;1:1–70. [PubMed]
  • Mansour SL, Thomas KR, Capecchi MR. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988 Nov 24;336(6197):348–352. [PubMed]
  • McMahon AP, Bradley A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell. 1990 Sep 21;62(6):1073–1085. [PubMed]
  • Moguilevsky N, Steens M, Thiriart C, Prieels JP, Thiry L, Bollen A. Lethal oxidative damage to human immunodeficiency virus by human recombinant myeloperoxidase. FEBS Lett. 1992 May 18;302(3):209–212. [PubMed]
  • Monaco AP. Molecular human genetics and the Duchenne/Becker muscular dystrophy gene. Mol Cell Biol Hum Dis Ser. 1993;3:1–11. [PubMed]
  • Nichols J, Evans EP, Smith AG. Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Development. 1990 Dec;110(4):1341–1348. [PubMed]
  • Nogae I, Johnston M. Isolation and characterization of the ZWF1 gene of Saccharomyces cerevisiae, encoding glucose-6-phosphate dehydrogenase. Gene. 1990 Dec 15;96(2):161–169. [PubMed]
  • Pandolfi PP, Roth ME, Karis A, Leonard MW, Dzierzak E, Grosveld FG, Engel JD, Lindenbaum MH. Targeted disruption of the GATA3 gene causes severe abnormalities in the nervous system and in fetal liver haematopoiesis. Nat Genet. 1995 Sep;11(1):40–44. [PubMed]
  • Rosenstraus M, Chasin LA. Isolation of mammalian cell mutants deficient in glucose-6-phosphate dehydrogenase activity: linkage to hypoxanthine phosphoribosyl transferase. Proc Natl Acad Sci U S A. 1975 Feb;72(2):493–497. [PMC free article] [PubMed]
  • Rudnicki MA, Braun T, Hinuma S, Jaenisch R. Inactivation of MyoD in mice leads to up-regulation of the myogenic HLH gene Myf-5 and results in apparently normal muscle development. Cell. 1992 Oct 30;71(3):383–390. [PubMed]
  • Smith AG, Heath JK, Donaldson DD, Wong GG, Moreau J, Stahl M, Rogers D. Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature. 1988 Dec 15;336(6200):688–690. [PubMed]
  • Thomas D, Cherest H, Surdin-Kerjan Y. Identification of the structural gene for glucose-6-phosphate dehydrogenase in yeast. Inactivation leads to a nutritional requirement for organic sulfur. EMBO J. 1991 Mar;10(3):547–553. [PMC free article] [PubMed]
  • Tinsley JM, Blake DJ, Pearce M, Knight AE, Kendrick-Jones J, Davies KE. Dystrophin and related proteins. Curr Opin Genet Dev. 1993 Jun;3(3):484–490. [PubMed]
  • Tsui LC. Mutations and sequence variations detected in the cystic fibrosis transmembrane conductance regulator (CFTR) gene: a report from the Cystic Fibrosis Genetic Analysis Consortium. Hum Mutat. 1992;1(3):197–203. [PubMed]
  • Vulliamy T, Mason P, Luzzatto L. The molecular basis of glucose-6-phosphate dehydrogenase deficiency. Trends Genet. 1992 Apr;8(4):138–143. [PubMed]
  • Vulliamy T, Beutler E, Luzzatto L. Variants of glucose-6-phosphate dehydrogenase are due to missense mutations spread throughout the coding region of the gene. Hum Mutat. 1993;2(3):159–167. [PubMed]
  • Wajntal A, DeMars R. A tetrazolium method for distinguishing between cultured human fibroblasts having eiter normal or deficient levels of glucose-6-phosphate dehydrogenase. Biochem Genet. 1967 Jun;1(1):61–64. [PubMed]
  • West JD. A genetically defined animal model of anembryonic pregnancy. Hum Reprod. 1993 Aug;8(8):1316–1323. [PubMed]
  • Williams RL, Hilton DJ, Pease S, Willson TA, Stewart CL, Gearing DP, Wagner EF, Metcalf D, Nicola NA, Gough NM. Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature. 1988 Dec 15;336(6200):684–687. [PubMed]
  • Winterbourn CC, Benatti U, De Flora A. Contributions of superoxide, hydrogen peroxide, and transition metal ions to auto-oxidation of the favism-inducing pyrimidine aglycone, divicine, and its reactions with haemoglobin. Biochem Pharmacol. 1986 Jun 15;35(12):2009–2015. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • Gene
    Gene links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • HomoloGene
    HomoloGene links
  • MedGen
    Related information in MedGen
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...