Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jul;131(1):18–29. doi: 10.1128/jb.131.1.18-29.1977

Control of Protein Synthesis in Escherichia coli: Analysis of an Energy Source Shift-Down

Knud Johnsen a, Søren Molin a,1, Olle Karlström a, Ole Maaløe a
PMCID: PMC235385  PMID: 326760

Abstract

The energy source shift-down described in the preceding paper (Molin et al., J. Bacteriol. 131: 7–17, 1977) was used to study the effects of shift-down on protein synthesis. The overall rate of protein synthesis was reduced immediately, and to the same extent, in stringent and relaxed strains. The primary effect of the shift was a slowing down of the polypeptide chain growth rate, a finding not previously reported. In stringent strains the normal, preshift rate was reestablished within 2 to 3 min, whereas in relaxed strains the chain growth rate remained low for about 20 min before slowly returning to the normal value, which was reestablished some 50 to 60 min after the shift. Throughout this transition, the stability of messenger ribonucleic acid (mRNA) remained unchanged in both strains. We interpret these findings as evidence of the more rapid reduction of the mRNA pool in the stringent strain after shift-down: we believe that very soon after the shift, the stringent strain reduces its pool of mRNA and with it the number of ribosomes engaged in protein synthesis. In this manner the number of active ribosomes is adjusted to the availability of energy and carbon. The relaxed strain cannot rapidly reduce its mRNA pool, which thus remains large enough to engage a near-preshift number of ribosomes during a prolonged period; as a consequence its ribosomes must work at a reduced rate. The possibility that ppGpp is involved in the control of mRNA production is discussed. After shift-down, the initial part of β-galactosidase (the auto-α fragment) was produced at a higher rate than complete β-galactosidase in the relaxed strain, as expected when translation is impeded.

Full text

PDF
18

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burgess R. R. Separation and characterization of the subunits of ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6168–6176. [PubMed] [Google Scholar]
  2. Cashel M., Gallant J. Two compounds implicated in the function of the RC gene of Escherichia coli. Nature. 1969 Mar 1;221(5183):838–841. doi: 10.1038/221838a0. [DOI] [PubMed] [Google Scholar]
  3. Coffman R. L., Norris T. E., Koch A. L. Chain elongation rate of messenger and polypeptides in slowly growing Escherichia coli. J Mol Biol. 1971 Aug 28;60(1):1–19. doi: 10.1016/0022-2836(71)90442-6. [DOI] [PubMed] [Google Scholar]
  4. Dalbow D. G., Young R. Synthesis time of beta-galactosidase in Escherichia coli B/r as a function of growth rate. Biochem J. 1975 Jul;150(1):13–20. doi: 10.1042/bj1500013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Errington L., Glass R. E., Hayward R. S., Scaife J. G. Structure and orientation of an RNA polymerase operon in Escherichia coli. Nature. 1974 Jun 7;249(457):519–522. doi: 10.1038/249519a0. [DOI] [PubMed] [Google Scholar]
  6. Fiil N. P., von Meyenburg K., Friesen J. D. Accumulation and turnover of guanosine tetraphosphate in Escherichia coli. J Mol Biol. 1972 Nov 28;71(3):769–783. doi: 10.1016/s0022-2836(72)80037-8. [DOI] [PubMed] [Google Scholar]
  7. Forchhammer J., Lindahl L. Growth rate of polypeptide chains as a function of the cell growth rate in a mutant of Escherichia coli 15. J Mol Biol. 1971 Feb 14;55(3):563–568. doi: 10.1016/0022-2836(71)90337-8. [DOI] [PubMed] [Google Scholar]
  8. Fowler A. V., Zabin I. The amino acid sequence of beta-galactosidase of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1507–1510. doi: 10.1073/pnas.74.4.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gausing K. Efficiency of protein and messenger RNA synthesis in bacteriophage T4-infected cells of Escherichia coli. J Mol Biol. 1972 Nov 28;71(3):529–545. doi: 10.1016/s0022-2836(72)80021-4. [DOI] [PubMed] [Google Scholar]
  10. Hall B., Gallant J. Defective translation in RC - cells. Nat New Biol. 1972 May 31;237(74):131–135. doi: 10.1038/newbio237131a0. [DOI] [PubMed] [Google Scholar]
  11. Hansen M. T., Bennett P. M., von Meyenburg K. Intracistronic polarity during dissociation of translation from transcription in Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):589–604. doi: 10.1016/0022-2836(73)90225-8. [DOI] [PubMed] [Google Scholar]
  12. Hansen M. T., Pato M. L., Molin S., Fill N. P., von Meyenburg K. Simple downshift and resulting lack of correlation between ppGpp pool size and ribonucleic acid accumulation. J Bacteriol. 1975 May;122(2):585–591. doi: 10.1128/jb.122.2.585-591.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Imamoto F. Diversity of regulation of genetic transcription. I. Effect of antibiotics which inhibit the process of translation on RNA metabolism in Escherichia coli. J Mol Biol. 1973 Feb 25;74(2):113–136. doi: 10.1016/0022-2836(73)90102-2. [DOI] [PubMed] [Google Scholar]
  14. Jacobson L. A., Baldassare J. C. Association of messenger ribonucleic acid with 70S monosomes from down-shifted Escherichia coli. J Bacteriol. 1976 Jul;127(1):637–643. doi: 10.1128/jb.127.1.637-643.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jacquet M., Kepes A. Initiation, elongation and inactivation of lac messenger RNA in Escherichia coli studied studied by measurement of its beta-galactosidase synthesizing capacity in vivo. J Mol Biol. 1971 Sep 28;60(3):453–472. doi: 10.1016/0022-2836(71)90181-1. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lin S., Zabin I. Beta-galactosidase. Rates of synthesis and degradation of incomplete chains. J Biol Chem. 1972 Apr 10;247(7):2205–2211. [PubMed] [Google Scholar]
  18. Matzura H., Molin S., Maaloe O. Sequential biosynthesis of the and ' subunits of the DNA-dependent RNA polymerase from Escherichia coli. J Mol Biol. 1971 Jul 14;59(1):17–25. doi: 10.1016/0022-2836(71)90410-4. [DOI] [PubMed] [Google Scholar]
  19. Midgley J. E. Turnover as a control of ribonucleic acid accumulation in bacteria undergoing stepdown. Biochem J. 1976 Feb 15;154(2):541–552. doi: 10.1042/bj1540541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Molin S., Von Meyenburg K., Maaloe O., Hansen M. T., Pato M. L. Control of ribosome synthesis in Escherichia coli: analysis of an energy source shift-down. J Bacteriol. 1977 Jul;131(1):7–17. doi: 10.1128/jb.131.1.7-17.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morrison S. L., Zipser D. Polypeptide products of nonsense mutations. I. Termination fragments from nonsense mutations in the Z gene of the lac operon of Escherichia coli. J Mol Biol. 1970 Jun 14;50(2):359–371. doi: 10.1016/0022-2836(70)90198-1. [DOI] [PubMed] [Google Scholar]
  22. Morse D. E., Guertin M. Regulation of mRNA utilization and degradation by amino-acid starvation. Nat New Biol. 1971 Aug 11;232(2):165–169. doi: 10.1038/newbio232165a0. [DOI] [PubMed] [Google Scholar]
  23. Morse D. E. Polarity induced by chloramphenicol and relief by suA. J Mol Biol. 1971 Jan 14;55(1):113–118. doi: 10.1016/0022-2836(71)90285-3. [DOI] [PubMed] [Google Scholar]
  24. Newton W. A., Beckwith J. R., Zipser D., Brenner S. Nonsense mutants and polarity in the lac operon of Escherichia coli. J Mol Biol. 1965 Nov;14(1):290–296. doi: 10.1016/s0022-2836(65)80250-9. [DOI] [PubMed] [Google Scholar]
  25. Pato M. L., Bennett P. M., von Meyenburg K. Messenger ribonucleic acid synthesis and degradation in Escherichia coli during inhibition of translation. J Bacteriol. 1973 Nov;116(2):710–718. doi: 10.1128/jb.116.2.710-718.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. STENT G. S., BRENNER S. A genetic locus for the regulation of ribonucleic acid synthesis. Proc Natl Acad Sci U S A. 1961 Dec 15;47:2005–2014. doi: 10.1073/pnas.47.12.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schleif R., Hess W., Finkelstein S., Ellis D. Induction kinetics of the L-arabinose operon of Escherichia coli. J Bacteriol. 1973 Jul;115(1):9–14. doi: 10.1128/jb.115.1.9-14.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sekiguchi M., Iida S. Mutants of Escherichia coli permeable to actinomycin. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2315–2320. doi: 10.1073/pnas.58.6.2315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Silverstone A. E., Arditti R. R., Magasanik B. Catabolite-insensitive revertants of lac promoter mutants. Proc Natl Acad Sci U S A. 1970 Jul;66(3):773–779. doi: 10.1073/pnas.66.3.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  31. Talkad V., Schneider E., Kennell D. Evidence for variable rates of ribosome movement in Escherichia coli. J Mol Biol. 1976 Jun 14;104(1):299–303. doi: 10.1016/0022-2836(76)90015-2. [DOI] [PubMed] [Google Scholar]
  32. Ullmann A., Jacob F., Monod J. Characterization by in vitro complementation of a peptide corresponding to an operator-proximal segment of the beta-galactosidase structural gene of Escherichia coli. J Mol Biol. 1967 Mar 14;24(2):339–343. doi: 10.1016/0022-2836(67)90341-5. [DOI] [PubMed] [Google Scholar]
  33. Ullmann A., Jacob F., Monod J. On the subunit structure of wild-type versus complemented beta-galactosidase of Escherichia coli. J Mol Biol. 1968 Feb 28;32(1):1–13. doi: 10.1016/0022-2836(68)90140-x. [DOI] [PubMed] [Google Scholar]
  34. Varmus H. E., Perlman R. L., Pastan I. Regulation of lac transcription in antibiotic-treated E. coli. Nat New Biol. 1971 Mar 10;230(10):41–44. doi: 10.1038/newbio230041a0. [DOI] [PubMed] [Google Scholar]
  35. Westover K. C., Jacobson L. A. Control of protein synthesis of Escherichia coli. I. Translation and functional inactivation of messenger ribonucleic acid after energy source shift-down. J Biol Chem. 1974 Oct 10;249(19):6272–6279. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES