Structural studies on colicin E3 and its immunity protein

  • M. Shoham
  • A. Djebli
Conference paper
Part of the NATO ASI Series book series (volume 65)

Abstract

Colicin E3 is a bacteriocin that penetrates sensitive cells and kills them by an enzymatic mode of action. It acts as a highly specific RNase that cleaves 16S ribosomal RNA at a position 49 bases in from the 3’end (Nomura et al. 1974). Although only a single phosphodiester bond is cleaved and the fragments stay attached to each other following the endonucleolytic attack, protein biosynthesis ceases completely and the infected cell dies. Colicin E3 acts in this specific manner only if all the components necessary for ribosome function are present. Thus, colicin E3 function requires the presence of the 50S subunit in addition to the 30S subunit as well as ribosomal proteins (Jakes & Zinder, 1974; Sidikaro & Nomura, 1974). Knowledge of the three-dimensional structure of colicin E3 will therefore not only provide an understanding of its mode of action but it is expected also to shed light on the structure of the ribosome at large in the vicinity of the cutting site. This region of the ribosome close to the interface between the small and large subunit is obviously critical for protein biosynthesis if a single nick in 16S RNA totally inactivates the ribosome.

Keywords

Crystallization Migration HPLC Mercury Citrate 

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References

  1. de Graaf FK, Klaasen-Boor P (1974) Purification and Characterization of the Cloacin DF13 Immunity Protein. FEBS Lett 40:293–296PubMedCrossRefGoogle Scholar
  2. Blundell TL, Johnson LN (1976) Protein Crystallography, pp. 224, Academic PressGoogle Scholar
  3. Frolow F, Shoham M (1990) Crystallization and Preliminary X-ray Investigation of Colicin E3 in Complex with Its Immunity Protein. J Biol Chem 265:10196–10197PubMedGoogle Scholar
  4. Herschman HR, Helinski DR (1967) Purification and Characterization of Colicin E2 and Colicin E3. J Biol Chem 242:5360–5368PubMedGoogle Scholar
  5. Jakes KS (1982) The mechanism of action of colicin E2, colicin E3 and cloacin DF13 in “Molecular action of toxins and viruses” (Cohen, P. and van Heyningen, S., eds), Elsevier Biomedical Press, Amsterdam-New York, OxfordGoogle Scholar
  6. Jakes KS, Zinder ND (1974) Highly Purified Colicin E3 Contains Immunity Protein Proc Natl Acad Sci USA 71:3380–3384PubMedCrossRefGoogle Scholar
  7. Jakes KS, Zinder N, Nomura M (1974) Purification and Properties of Colicin E3 Immunity Protein. J.Biol Chem 249:438–444PubMedGoogle Scholar
  8. Lau C, Richards FM (1976) Proteolytic and Chemical Modification of Colicin E3 Activity. Biochemistry 15:3856–3863PubMedCrossRefGoogle Scholar
  9. Levinson BL, Pickover CA, Richards FM (1983) Dimerization of Colicin E3* in the Absence of Immunity Protein. J Biol Chem 258:10967–10972PubMedGoogle Scholar
  10. Masaki H, Ohta T (1985) Colicin E3 and Its Immunity Genes. J Mol Biol 182:217–227PubMedCrossRefGoogle Scholar
  11. Mochitate K, Suzuki K, Imahori K (1981) Amino Acid Sequence of Immunity Protein (B Subunit) of Colicin E3. J.Biochem (Tokyo) 89:1609–1618PubMedGoogle Scholar
  12. Nomura M, Sidikaro J, Jakes K, Zinder P(1974) In: Nomura M, Tissiers A, Lenzyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, Cold Spring Harbor New York, 805–814Google Scholar
  13. Ohno-Iwashita Y, Imahori K (1977) Colicin E3 Induced Cleavage of 16S rRNA of Isolated 30S Ribosomal Subunits. J Biochem (Tokyo) 82:919–922Google Scholar
  14. Ohno S, Ohno-Iwashita Y, Suzuki K, Imahori K (1977) Purification and characterization of active component and active fragment of colicin E3. J Biochem (Tokyo) 82:1045–1053PubMedGoogle Scholar
  15. Ohno S, Imahori K (1978) Colicin E3 is an Endonuclease. J Biochem (Tokyo) 84:1637–1640PubMedGoogle Scholar
  16. Sabet SF, Schnaitman CA (1973) Purification and Properties of the Colicin E3 Receptor of Escherichia Coli. J Biol Chem 248:1797–1806PubMedGoogle Scholar
  17. Shoham M, Levinson BL, Richards FM (1984) Crystallization and Preliminary X-ray Diffraction Studies of Colicin E3 Immunity Protein. J Mol Biol 177:563–565PubMedCrossRefGoogle Scholar
  18. Sidikaro J, Nomura M (1974) A Protein from E3-Colicinogenic Cells That Accounts for Their Immunity to Colicin E3. J Biol Chem 249:445–453PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1992

Authors and Affiliations

  • M. Shoham
    • 1
  • A. Djebli
    • 1
  1. 1.Department of Biochemistry School of MedicineCase Western Reserve UniversityClevelandUSA

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