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Biochem J. Apr 15, 2004; 379(Pt 2): 243–251.
PMCID: PMC1224093

Inosine 5'-monophosphate dehydrogenase binds nucleic acids in vitro and in vivo.

Abstract

Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in the de novo biosynthesis of guanine nucleotides. In addition to the catalytic domain, IMPDH contains a subdomain of unknown function composed of two cystathione beta-synthase domains. Our results, using three different assays, show that IMPDHs from Tritrichomonas foetus, Escherichia coli, and both human isoforms bind single-stranded nucleic acids with nanomolar affinity via the subdomain. Approx. 100 nucleotides are bound per IMPDH tetramer. Deletion of the subdomain decreases affinity 10-fold and decreases site size to 60 nucleotides, whereas substitution of conserved Arg/Lys residues in the subdomain with Glu decreases affinity by 20-fold. IMPDH is found in the nucleus of human cells, as might be expected for a nucleic-acid-binding protein. Lastly, immunoprecipitation experiments show that IMPDH binds both RNA and DNA in vivo. These experiments indicate that IMPDH has a previously unappreciated role in replication, transcription or translation that is mediated by the subdomain.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Weber G, Nakamura H, Natsumeda Y, Szekeres T, Nagai M. Regulation of GTP biosynthesis. Adv Enzyme Regul. 1992;32:57–69. [PubMed]
  • Allison AC, Almquist SJ, Muller CD, Eugui EM. In vitro immunosuppressive effects of mycophenolic acid and an ester pro-drug, RS-61443. Transplant Proc. 1991 Apr;23(2 Suppl 2):10–14. [PubMed]
  • De Clercq E. Antiviral agents: characteristic activity spectrum depending on the molecular target with which they interact. Adv Virus Res. 1993;42:1–55. [PubMed]
  • Robins RK, Revankar GR, McKernan PA, Murray BK, Kirsi JJ, North JA. The importance of IMP dehydrogenase inhibition in the broad spectrum antiviral activity of ribavirin and selenazofurin. Adv Enzyme Regul. 1985;24:29–43. [PubMed]
  • Weber G. Biochemical strategy of cancer cells and the design of chemotherapy: G. H. A. Clowes Memorial Lecture. Cancer Res. 1983 Aug;43(8):3466–3492. [PubMed]
  • Sintchak MD, Fleming MA, Futer O, Raybuck SA, Chambers SP, Caron PR, Murcko MA, Wilson KP. Structure and mechanism of inosine monophosphate dehydrogenase in complex with the immunosuppressant mycophenolic acid. Cell. 1996 Jun 14;85(6):921–930. [PubMed]
  • Nimmesgern E, Black J, Futer O, Fulghum JR, Chambers SP, Brummel CL, Raybuck SA, Sintchak MD. Biochemical analysis of the modular enzyme inosine 5'-monophosphate dehydrogenase. Protein Expr Purif. 1999 Nov;17(2):282–289. [PubMed]
  • Gan Lu, Petsko Gregory A, Hedstrom Lizbeth. Crystal structure of a ternary complex of Tritrichomonas foetus inosine 5'-monophosphate dehydrogenase: NAD+ orients the active site loop for catalysis. Biochemistry. 2002 Nov 5;41(44):13309–13317. [PubMed]
  • Bowne Sara J, Sullivan Lori S, Blanton Susan H, Cepko Constance L, Blackshaw Seth, Birch David G, Hughbanks-Wheaton Dianna, Heckenlively John R, Daiger Stephen P. Mutations in the inosine monophosphate dehydrogenase 1 gene (IMPDH1) cause the RP10 form of autosomal dominant retinitis pigmentosa. Hum Mol Genet. 2002 Mar 1;11(5):559–568. [PMC free article] [PubMed]
  • Kennan Avril, Aherne Aileen, Palfi Arpad, Humphries Marian, McKee Alex, Stitt Alan, Simpson David A C, Demtroder Karin, Orntoft Torben, Ayuso Carmen, et al. Identification of an IMPDH1 mutation in autosomal dominant retinitis pigmentosa (RP10) revealed following comparative microarray analysis of transcripts derived from retinas of wild-type and Rho(-/-) mice. Hum Mol Genet. 2002 Mar 1;11(5):547–557. [PubMed]
  • Cornuel Jean François, Moraillon Anne, Guéron Maurice. Participation of yeast inosine 5'-monophosphate dehydrogenase in an in vitro complex with a fragment of the C-rich telomeric strand. Biochimie. 2002 Apr;84(4):279–289. [PubMed]
  • Digits JA, Hedstrom L. Kinetic mechanism of Tritrichomonas foetus inosine 5'-monophosphate dehydrogenase. Biochemistry. 1999 Feb 23;38(8):2295–2306. [PubMed]
  • Farazi T, Leichman J, Harris T, Cahoon M, Hedstrom L. Isolation and characterization of mycophenolic acid-resistant mutants of inosine-5'-monophosphate dehydrogenase. J Biol Chem. 1997 Jan 10;272(2):961–965. [PubMed]
  • Kerr KM, Hedstrom L. The roles of conserved carboxylate residues in IMP dehydrogenase and identification of a transition state analog. Biochemistry. 1997 Oct 28;36(43):13365–13373. [PubMed]
  • Bock LC, Griffin LC, Latham JA, Vermaas EH, Toole JJ. Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature. 1992 Feb 6;355(6360):564–566. [PubMed]
  • Singh R, Green MR. Sequence-specific binding of transfer RNA by glyceraldehyde-3-phosphate dehydrogenase. Science. 1993 Jan 15;259(5093):365–368. [PubMed]
  • Griffoni C, Laktionov PP, Rykova EY, Spisni E, Riccio M, Santi S, Bryksin A, Volodko N, Kraft R, Vlassov V, et al. The Rossmann fold of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a nuclear docking site for antisense oligonucleotides containing a TAAAT motif. Biochim Biophys Acta. 2001 Jan 15;1530(1):32–46. [PubMed]
  • Lohman TM, Ferrari ME. Escherichia coli single-stranded DNA-binding protein: multiple DNA-binding modes and cooperativities. Annu Rev Biochem. 1994;63:527–570. [PubMed]
  • Bujalowski W, Overman LB, Lohman TM. Binding mode transitions of Escherichia coli single strand binding protein-single-stranded DNA complexes. Cation, anion, pH, and binding density effects. J Biol Chem. 1988 Apr 5;263(10):4629–4640. [PubMed]
  • Cokol M, Nair R, Rost B. Finding nuclear localization signals. EMBO Rep. 2000 Nov;1(5):411–415. [PMC free article] [PubMed]
  • Nakai K, Horton P. PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci. 1999 Jan;24(1):34–36. [PubMed]
  • Chook YM, Blobel G. Karyopherins and nuclear import. Curr Opin Struct Biol. 2001 Dec;11(6):703–715. [PubMed]
  • Morgenegg G, Winkler GC, Hübscher U, Heizmann CW, Mous J, Kuenzle CC. Glyceraldehyde-3-phosphate dehydrogenase is a nonhistone protein and a possible activator of transcription in neurons. J Neurochem. 1986 Jul;47(1):54–62. [PubMed]
  • De BP, Gupta S, Zhao H, Drazba JA, Banerjee AK. Specific interaction in vitro and in vivo of glyceraldehyde-3-phosphate dehydrogenase and LA protein with cis-acting RNAs of human parainfluenza virus type 3. J Biol Chem. 1996 Oct 4;271(40):24728–24735. [PubMed]
  • Schultz DE, Hardin CC, Lemon SM. Specific interaction of glyceraldehyde 3-phosphate dehydrogenase with the 5'-nontranslated RNA of hepatitis A virus. J Biol Chem. 1996 Jun 14;271(24):14134–14142. [PubMed]
  • Zang WQ, Fieno AM, Grant RA, Yen TS. Identification of glyceraldehyde-3-phosphate dehydrogenase as a cellular protein that binds to the hepatitis B virus posttranscriptional regulatory element. Virology. 1998 Aug 15;248(1):46–52. [PubMed]
  • Nagy E, Rigby WF. Glyceraldehyde-3-phosphate dehydrogenase selectively binds AU-rich RNA in the NAD(+)-binding region (Rossmann fold). J Biol Chem. 1995 Feb 10;270(6):2755–2763. [PubMed]
  • Sioud M, Jespersen L. Enhancement of hammerhead ribozyme catalysis by glyceraldehyde-3-phosphate dehydrogenase. J Mol Biol. 1996 Apr 12;257(4):775–789. [PubMed]
  • Meyer-Siegler K, Mauro DJ, Seal G, Wurzer J, deRiel JK, Sirover MA. A human nuclear uracil DNA glycosylase is the 37-kDa subunit of glyceraldehyde-3-phosphate dehydrogenase. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8460–8464. [PMC free article] [PubMed]
  • Lin SS, Chang SC, Wang YH, Sun CY, Chang MF. Specific interaction between the hepatitis delta virus RNA and glyceraldehyde 3-phosphate dehydrogenase: an enhancement on ribozyme catalysis. Virology. 2000 May 25;271(1):46–57. [PubMed]
  • Evguenieva-Hackenberg Elena, Schiltz Emile, Klug Gabriele. Dehydrogenases from all three domains of life cleave RNA. J Biol Chem. 2002 Nov 29;277(48):46145–46150. [PubMed]
  • Ishitani R, Tanaka M, Sunaga K, Katsube N, Chuang DM. Nuclear localization of overexpressed glyceraldehyde-3-phosphate dehydrogenase in cultured cerebellar neurons undergoing apoptosis. Mol Pharmacol. 1998 Apr;53(4):701–707. [PubMed]
  • Saunders PA, Chen RW, Chuang DM. Nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase isoforms during neuronal apoptosis. J Neurochem. 1999 Mar;72(3):925–932. [PubMed]
  • Dastoor Z, Dreyer JL. Potential role of nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase in apoptosis and oxidative stress. J Cell Sci. 2001 May;114(Pt 9):1643–1653. [PubMed]
  • Sawa A, Khan AA, Hester LD, Snyder SH. Glyceraldehyde-3-phosphate dehydrogenase: nuclear translocation participates in neuronal and nonneuronal cell death. Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11669–11674. [PMC free article] [PubMed]
  • Choudhary S, De BP, Banerjee AK. Specific phosphorylated forms of glyceraldehyde 3-phosphate dehydrogenase associate with human parainfluenza virus type 3 and inhibit viral transcription in vitro. J Virol. 2000 Apr;74(8):3634–3641. [PMC free article] [PubMed]
  • Bateman A. The structure of a domain common to archaebacteria and the homocystinuria disease protein. Trends Biochem Sci. 1997 Jan;22(1):12–13. [PubMed]
  • Ponting CP. CBS domains in CIC chloride channels implicated in myotonia and nephrolithiasis (kidney stones). J Mol Med (Berl) 1997 Mar;75(3):160–163. [PubMed]
  • Blair E, Redwood C, Ashrafian H, Oliveira M, Broxholme J, Kerr B, Salmon A, Ostman-Smith I, Watkins H. Mutations in the gamma(2) subunit of AMP-activated protein kinase cause familial hypertrophic cardiomyopathy: evidence for the central role of energy compromise in disease pathogenesis. Hum Mol Genet. 2001 May 15;10(11):1215–1220. [PubMed]
  • Farrar G Jane, Kenna Paul F, Humphries Peter. On the genetics of retinitis pigmentosa and on mutation-independent approaches to therapeutic intervention. EMBO J. 2002 Mar 1;21(5):857–864. [PMC free article] [PubMed]
  • Chu E, Allegra CJ. The role of thymidylate synthase in cellular regulation. Adv Enzyme Regul. 1996;36:143–163. [PubMed]
  • Zhang Kai, Rathod Pradipsinh K. Divergent regulation of dihydrofolate reductase between malaria parasite and human host. Science. 2002 Apr 19;296(5567):545–547. [PMC free article] [PubMed]
  • Allison AC, Eugui EM. Mycophenolate mofetil and its mechanisms of action. Immunopharmacology. 2000 May;47(2-3):85–118. [PubMed]
  • Shaw RJ, Wilson JL, Smith KT, Reines D. Regulation of an IMP dehydrogenase gene and its overexpression in drug-sensitive transcription elongation mutants of yeast. J Biol Chem. 2001 Aug 31;276(35):32905–32916. [PMC free article] [PubMed]
  • Uetz P, Giot L, Cagney G, Mansfield TA, Judson RS, Knight JR, Lockshon D, Narayan V, Srinivasan M, Pochart P, et al. A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature. 2000 Feb 10;403(6770):623–627. [PubMed]
  • Ito T, Chiba T, Ozawa R, Yoshida M, Hattori M, Sakaki Y. A comprehensive two-hybrid analysis to explore the yeast protein interactome. Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4569–4574. [PMC free article] [PubMed]
  • Ho Yuen, Gruhler Albrecht, Heilbut Adrian, Bader Gary D, Moore Lynda, Adams Sally-Lin, Millar Anna, Taylor Paul, Bennett Keiryn, Boutilier Kelly, et al. Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry. Nature. 2002 Jan 10;415(6868):180–183. [PubMed]
  • Lohman TM, Mascotti DP. Thermodynamics of ligand-nucleic acid interactions. Methods Enzymol. 1992;212:400–424. [PubMed]

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