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Characterization of Genes for Polyamine Modulon

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Polyamines

Part of the book series: Methods in Molecular Biology ((MIMB,volume 720))

Abstract

Polyamines are essential for normal cell growth and exist mainly as RNA-polyamine complexes in cells. Thus, effects of polyamines on protein synthesis have been studied. It was found that several kinds of protein synthesis, which are important for cell growth, were enhanced by polyamines at the level of translation. We proposed that a group of genes whose expression is enhanced by polyamines at the level of translation be referred to as a “polyamine modulon.” In Escherichia coli, most members of the polyamine modulon thus far identified were transcription factors. These transcription factors enhanced the synthesis of several kinds of mRNA and tRNA, and also rRNA. In this way, polyamines enhanced growth of E. coli. We also succeeded in identifying three kinds of “polyamine modulon” in mammalian cells. One of the mechanisms of polyamine stimulation at the molecular level was due to the stabilization of the bulged-out region of double-stranded RNA in mRNA. The procedures used to identify components of the polyamine modulon are described in this chapter.

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References

  1. Cohen SS (1998) A guide to polyamines. Oxford University Press, New York, pp 1–543

    Google Scholar 

  2. Igarashi K, Kashiwagi K (2000) Polyamines: mysterious modulators of cellular functions. Biochem Biophys Res Commun 271:559–564

    Article  PubMed  CAS  Google Scholar 

  3. Watanabe S, Kusama-Eguchi K, Kobayashi H, Igarashi K (1991) Estimation of polyamine binding to macromolecules and ATP in bovine lymphocytes and rat liver. J Biol Chem 266:20803–20809

    PubMed  CAS  Google Scholar 

  4. Miyamoto S, Kashiwagi K, Ito K, Watanabe S, Igarashi K (1993) Estimation of polyamine distribution and polyamine stimulation of protein synthesis in Escherichia coli. Arch Biochem Biophys 300:63–68

    Article  PubMed  CAS  Google Scholar 

  5. Cunningham-Rundles S, Maas WK (1975) Isolation, characterization, and mapping of Escherichia coli mutants blocked in the synthesis of ornithine decarboxylase. J Bacteriol 124:791–799

    PubMed  CAS  Google Scholar 

  6. Linderoth N, Morris DR (1983) Structural specificity of the triamines sym-homospermidine and aminopropylcadaverine in stimulating growth of spermidine auxotrophs of Escherichia coli. Biochem Biophys Res Commun 117:616–622

    Article  PubMed  CAS  Google Scholar 

  7. Hafner EW, Tabor CW, Tabor H (1979) Mutants of Escherichia coli that do not contain 1, 4-diaminobutane (putrescine) or spermidine. J Biol Chem 254:12419–12426

    PubMed  CAS  Google Scholar 

  8. Kashiwagi K, Miyaji A, Ikeda S, Tobe T, Sasakawa C, Igarashi K (1992) Increase of sensitivity to aminoglycoside antibiotics by polyamine-induced protein (oligopeptide-binding protein) in Escherichia coli. J Bacteriol 174:4331–4337

    PubMed  CAS  Google Scholar 

  9. Kashiwagi K, Watanabe R, Igarashi K (1994) Involvement of ribonuclease III in the enhancement of expression of the speF-potE operon encoding inducible ornithine decarboxylase and polyamine transport protein. Biochem Biophys Res Commun 200:591–597

    Article  PubMed  CAS  Google Scholar 

  10. Kashiwagi K, Yamaguchi Y, Sakai Y, Kobayashi H, Igarashi K (1990) Identification of the polyamine-induced protein as a periplasmic oligopeptide binding protein. J Biol Chem 265:8387–8391

    PubMed  CAS  Google Scholar 

  11. Igarashi K, Saisho T, Yuguchi M, Kashiwagi K (1997) Molecular mechanism of polyamine stimulation of the synthesis of oligopeptide-binding protein. J Biol Chem 272:4058–4064

    Article  PubMed  CAS  Google Scholar 

  12. Higashi K, Terui Y, Suganami A, Tamura Y, Nishimura K, Kashiwagi K, Igarashi K (2008) Selective structural change by spermidine in the bulged-out region of double-stranded RNA and its effect on RNA function. J Biol Chem 283:32989–32994

    Article  PubMed  CAS  Google Scholar 

  13. Shapira SK, Chou J, Richaud FV, Casadaban MJ (1983) New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of β-galactosidase. Gene 25:71–82

    Article  PubMed  CAS  Google Scholar 

  14. Masaki T, Tanabe M, Nakamura K, Soejima M (1981) Studies on a new proteolytic enzyme from Achromobacter lyticus M497-1. I. Purification and some enzymatic properties. Biochim Biophys Acta 660:44–50

    PubMed  CAS  Google Scholar 

  15. Emory SA, Belasco JG (1990) The ompA 5’ untranslated RNA segment functions in Escherichia coli as a growth-rate-regulated mRNA stabilizer whose activity is unrelated to translational efficiency. J Bacteriol 172:4472–4481

    PubMed  CAS  Google Scholar 

  16. Sambrook J, Fritsch EF, Maniatis T (2001) Extraction, purification, and analysis of mRNA from eukaryotic cells, Chapter 7. In: Sambrook J, Russell DW (eds) Molecular cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  18. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685

    Article  CAS  Google Scholar 

  19. Nielsen PJ, Manchester KL, Towbin H, Gordon J, Thomas G (1982) The phosphorylation of ribosomal protein S6 in rat tissues following cycloheximide injection, in diabetes, and after denervation of diaphragm. A simple immunological determination of the extent of S6 phosphorylation on protein blots. J Biol Chem 257:12316–12321

    PubMed  CAS  Google Scholar 

  20. Philipson L, Andersson P, Olshevsky U, Weinberg R, Baltimore D, Gesteland R (1978) Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA. Cell 13:189–199

    Article  PubMed  CAS  Google Scholar 

  21. Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59

    Article  PubMed  CAS  Google Scholar 

  22. Ayusawa D, Iwata K, Seno T (1981) Alteration of ribonucleotide reductase in aphidicolin-resistant mutants of mouse FM3A cells with associated resistance to arabinosyladenine and arabinosylcytosine. Somatic Cell Genet 7:27–42

    Article  PubMed  CAS  Google Scholar 

  23. Yoshida M, Meksuriyen D, Kashiwagi K, Kawai G, Igarashi K (1999) Polyamine stimulation of the synthesis of oligopeptide-binding protein (OppA). Involvement of a structural change of the Shine-Dalgarno sequence and the initiation codon AUG in OppA mRNA. J Biol Chem 274:22723–22728

    Article  PubMed  CAS  Google Scholar 

  24. Yoshida M, Kashiwagi K, Shigemasa A, Taniguchi S, Yamamoto K, Makinoshima H, Ishihama A, Igarashi K (2004) A unifying model for the role of polyamines in bacterial cell growth, the polyamine modulon. J Biol Chem 279:46008–46013

    Article  PubMed  CAS  Google Scholar 

  25. Terui Y, Higashi K, Taniguchi S, Shigemasa A, Nishimura K, Yamamoto K, Kashiwagi K, Ishihama A, Igarashi K (2007) Enhancement of the synthesis of RpoN, Cra and H-NS by polyamines at the level of translation in Escherichia coli cultured with glucose and glutamate. J Bacteriol 189:2359–2368

    Article  PubMed  CAS  Google Scholar 

  26. Terui Y, Higashi K, Tabei Y, Tomitori H, Yamamoto K, Ishihama A, Igarashi K, Kashiwagi K (2009) Enhancement of the synthesis of RpoE and StpA by polyamines at the level of translation in Escherichia coli under heat shock conditions. J Bacteriol 191:5348–5357

    Article  PubMed  CAS  Google Scholar 

  27. Yoshida M, Kashiwagi K, Kawai G, Ishihama A, Igarashi K (2001) Polyamine enhancement of the synthesis of adenylate cyclase at the translational level and cosequential stimulation of the synthesis of the RNA polymerase σ28 subunit. J Biol Chem 276:16289–16295

    Article  PubMed  CAS  Google Scholar 

  28. Yoshida M, Kashiwagi K, Kawai G, Ishihama A, Igarashi K (2002) Polyamines enhance synthesis of the RNA polymerase σ38 Subunit by suppression of an amber termination codon in the open reading frame. J Biol Chem 277:37139–37146

    Article  PubMed  CAS  Google Scholar 

  29. Higashi K, Kashiwagi K, Taniguchi S, Terui Y, Yamamoto K, Ishihama A, Igarashi K (2006) Enhancement of +1 frameshift by polyamines during translation of polypeptide release factor 2 in Escherichia coli. J Biol Chem 281:9527–9537

    Article  PubMed  CAS  Google Scholar 

  30. Higashi K, Terui Y, Inomata E, Katagiri D, Nomura Y, Someya T, Nishimura K, Kashiwagi K, Kawai G, Igarashi K (2008) Selective structural change of bulged-out region of double-stranded RNA containing bulged nucleotides by spermidine. Biochem Biophys Res Commun 370:572–577

    Article  PubMed  CAS  Google Scholar 

  31. Nishimura K, Murozumi K, Shirahata A, Park MH, Kashiwagi K, Igarashi K (2005) Independent roles of eIF5A and polyamines in cell proliferation. Biochem J 385:779–785

    Article  PubMed  CAS  Google Scholar 

  32. Nishimura K, Nakatsu F, Kashiwagi K, Ohno H, Saito T, Igarashi K (2002) Essential role of S-adenosylmethionine decarboxylase in mouse embryonic development. Genes Cells 7:41–47

    Article  PubMed  CAS  Google Scholar 

  33. Nishimura K, Okudaira H, Ochiai E, Higashi K, Kaneko M, Ishii I, Nishimura T, Dohmae N, Kashiwagi K, Igarashi K (2009) Identification of proteins whose synthesis is preferentially enhanced by polyamines at the level of translation in mammalian cells. Int J Biochem Cell Biol 41:2251–2261

    Article  PubMed  CAS  Google Scholar 

  34. Chappell SA, Dresios J, Edelman GM, Mauro VP (2006) Ribosomal shunting mediated by a translational enhancer element that base pairs to 18S rRNA. Proc Natl Acad Sci USA 103:9488–9493

    Article  PubMed  CAS  Google Scholar 

  35. Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Drs. K. Williams and A. J. Michael for critical reading of the manuscript prior to submission. This study was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan.

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Correspondence to Kazuei Igarashi .

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Igarashi, K., Kashiwagi, K. (2011). Characterization of Genes for Polyamine Modulon. In: Pegg, A., Casero, Jr., R. (eds) Polyamines. Methods in Molecular Biology, vol 720. Humana Press. https://doi.org/10.1007/978-1-61779-034-8_3

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  • DOI: https://doi.org/10.1007/978-1-61779-034-8_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-033-1

  • Online ISBN: 978-1-61779-034-8

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