Current Microbiology

, Volume 21, Issue 2, pp 131–137 | Cite as

Genetic regulation of glycogen biosynthesis inEscherichia coli: In vivo effects of the catabolite repression and stringent response systems inglg gene expression

  • Tony Romeo
  • Jill Black
  • Jack Preiss
Article

Abstract

The synthesis of two of theEscherichia coli glycogen biosynthetic enzymes, ADPglucose pyrophosphorylase (glgC) and glycogen synthase (glgA) was activated by the addition of 5 mM cyclic AMP (cAMP) to maxicells; synthesis of glycogen branching enzyme (glgB) was unaffected.β-Galactosidase activity expressed from a gene fusion,φ(glgC-lacZ), was approximately five-fold higher in acya+ versus an isogeniccya strain ofE. coli. Addition of cAMP restoredβ-galactosidase in thecya strain. The expression ofφ(glgC‘−’lacZ) encodedβ-galactosidase activity in a series ofspoT mutants exhibited an apparent exponential relationship to intracellular guanosine 5′-diphosphate 3′-diphosphate (ppGpp) levels. These results provide evidence for the control of glycogen biosynthesis in vivo by cAMP and ppGpp at the level of gene expression, and identify a region of DNA required for the control. Theφ(glgC‘−’lacZ) encodedβ-galactosidase activity was also elevated three-to five-fold in strain AC70R1, which contains a transacting mutation (glgQ) that affects the levels of the glycogen biosynthetic enzymes andglgC transcripts.

Keywords

Enzyme Gene Expression Response System Diphosphate Gene Fusion 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature Cited

  1. 1.
    Baracchini E, Bremer H (1988) Stringent and growth control of rRNA synthesis inEscherichia coli are both mediated by ppGPp. J Biol Chem 263:2597–2602PubMedGoogle Scholar
  2. 2.
    Cashel M, Rudd KE (1987) The stringent response. In: Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger E (eds)Escherichia coli andSalmonella typhimurium: cellular and molecular biology, Vol. 1. Washington, DC: American Society for MicrobiologyGoogle Scholar
  3. 3.
    Guerinot L, Chelm BK (1984) Isolation and expression of theBradyrhizobium japonicum adenylate cyclase gene (cya) inEscherichia coli. J Bacteriol 159:1068–1071PubMedGoogle Scholar
  4. 4.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  5. 5.
    Lorence MC, Alcorn JL, Rupter CS (1985) Construction of an improved maxicell strain for the identification of recombinant plasmid encoded proteins. In: Hollaender A (ed) Plasmids in bacteria, Basic life sciences, vol.30. pp 955Google Scholar
  6. 6.
    Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor LaboratoryGoogle Scholar
  7. 7.
    Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor, NY: Cold Spring Harbor LaboratoryGoogle Scholar
  8. 8.
    Okita TW, Rodriguez RL, Preiss J (1981) Biosynthesis of bacterial glycogen. Cloning of the glycogen biosynthetic enzyme structural genes. J Biol Chem 256:6944–6952PubMedGoogle Scholar
  9. 9.
    Preiss J (1984) Bacterial glycogen biosynthesis and its regulation. Annu Rev Microbiol 38:410–458CrossRefGoogle Scholar
  10. 10.
    Preiss J, Romeo T (1989) Physiology, biochemistry and genetics of bacterial glycogen synthesis. Adv Microb Physiol 30:183–238PubMedGoogle Scholar
  11. 11.
    Preiss J, Walsh DA (1981) The comparative biochemistry of glycogen and starch. Biol Carbohydr 1:199–314Google Scholar
  12. 12.
    Preiss J, Yung S-G, Baecker PA (1983) Regulation of bacterial glycogen biosynthesis. Mol Cell Biochem 57:61–80CrossRefPubMedGoogle Scholar
  13. 13.
    Primakoff P (1981) In vivo role of therelA+ gene in the regulation of thelac operon. J Bacteriol 145:410–416PubMedGoogle Scholar
  14. 14.
    Primakoff P, Artz SW (1979) Positive control oflac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate. Proc Natl Acad Sci USA 76:1726–1730PubMedGoogle Scholar
  15. 15.
    Romeo T, Preiss J (1989) Genetic regulation of glycogen biosynthesis inEscherichia coli: in vitro effects of cyclic AMP and guanosine 5′-diphosphate 3′-diphosphate and analysis of in vivo transcripts. J Bacteriol 171:2773–2782PubMedGoogle Scholar
  16. 16.
    Romeo T, Kumar A, Preiss J (1988) Analysis of theEscherichia coli glycogen gene cluster suggests that catabolic enzymes are encoded among the biosynthetic genes. Gene 70:363–376CrossRefPubMedGoogle Scholar
  17. 17.
    Sancar A, Hack AM, Rupp DW (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693PubMedGoogle Scholar
  18. 18.
    Sarubbi E, Rudd KE, Cashel M (1988) Basal ppGpp level adjustment shown by newspoT mutants affect steady state growth rates and rrnA ribosomal promoter regulation inEscherichia coli. Mol Gen Genet 213:214–222CrossRefPubMedGoogle Scholar
  19. 19.
    Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor, New York, Cold Spring Harbor LaboratoryGoogle Scholar
  20. 20.
    Urbanowski J, Leung P, Weissbach H, Preiss J (1983) The in vitro expression of the gene forEscherichia coli ADP glucose pyrophosphorylase is stimulated by cyclic AMP and cyclic AMP receptor protein. J Biol Chem 258:2782–2784PubMedGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1990

Authors and Affiliations

  • Tony Romeo
    • 1
  • Jill Black
    • 1
  • Jack Preiss
    • 1
  1. 1.Department of BiochemistryMichigan State UniversityEast LansingUSA

Personalised recommendations