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. 1977 Dec;74(12):5387–5391. doi: 10.1073/pnas.74.12.5387

mRNA-dependent in vitro synthesis of ribosomal proteins L12 and L10 and elongation factor Tu.

F Chu, P Caldwell, H Weissbach, N Brot
PMCID: PMC431730  PMID: 341155

Abstract

RNA extracted from growing Escherichia coli can direct the in vitro synthesis of ribosomal proteins L12 and L10 and elongation factor Tu when an E. coli system is used. The synthesized L12 can be bound to L12-depleted ribosomes and the synthesized elongation factor Tu can form complexes with both elongation factor Ts and GDP. Guanosine 5'-diphosphate 3'-diphosphate has no effect on the synthesis of these proteins from an RNA template but inhibits their synthesis when a DNA template is used.

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Selected References

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

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Brot N., Marcel R., Yamasaki E., Weissbach H. Further studies on the role of 50 S ribosomal proteins in protein synthesis. J Biol Chem. 1973 Oct 25;248(20):6952–6956. [PubMed] [Google Scholar]
  3. Chu F., Caldwell P., Samuels M., Weissbach H., Brot N. DNA-dependent in vitro synthesis of Escherichia coli ribosomal protein L10 and the formation of an L10L12 complex. Biochem Biophys Res Commun. 1976 May 23;76(2):593–601. doi: 10.1016/0006-291x(77)90765-3. [DOI] [PubMed] [Google Scholar]
  4. Chu F., Kung H. F., Caldwell P., Weissbach H., Brot N. DNA dependent synthesis of protein L12 from escherichia coli ribosomes, in vitro. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3156–3159. doi: 10.1073/pnas.73.9.3156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chu F., Miller D. L., Schulz T., Weissbach H., Brot N. DNA-directed in vitro synthesis of elongation factor Tu. Biochem Biophys Res Commun. 1976 Dec 20;73(4):917–927. doi: 10.1016/0006-291x(76)90209-6. [DOI] [PubMed] [Google Scholar]
  6. Dennis P. P., Nomura M. Stringent control of ribosomal protein gene expression in Escherichia coli. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3819–3823. doi: 10.1073/pnas.71.10.3819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dennis P. P., Nomura M. Stringent control of the transcriptional activities of ribosomal protein genes in E. coli. Nature. 1975 Jun 5;255(5508):460–465. doi: 10.1038/255460a0. [DOI] [PubMed] [Google Scholar]
  8. Deusser E., Weber J., Maschler R., Stöffler G., Wittmann H. G. Structural and functional evidence for a repeated 50S subunit ribosomal protein. Nat New Biol. 1973 Mar 14;242(115):47–49. doi: 10.1038/newbio242047a0. [DOI] [PubMed] [Google Scholar]
  9. Dohan F. C., Jr, Rubman R. H., Torriani A. In vitro synthesis of Escherichia coli alkaline phosphatase monomers. J Mol Biol. 1971 Jun 14;58(2):469–479. doi: 10.1016/0022-2836(71)90364-0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Furano A. V. Content of elongation factor Tu in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4780–4784. doi: 10.1073/pnas.72.12.4780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Furano A. V., Wittel F. P. Syntheses of elongation factors Tu and G are under stringent control in Escherichia coli. J Biol Chem. 1976 Feb 10;251(3):898–901. [PubMed] [Google Scholar]
  13. Hachmann J., Miller D. L., Weissbach H. Purification of factor Ts: studies on the formation and stability of nucleotide complexes containing transfer factor Tu. Arch Biochem Biophys. 1971 Dec;147(2):457–466. doi: 10.1016/0003-9861(71)90401-2. [DOI] [PubMed] [Google Scholar]
  14. Hamel E., Koka M., Nakamoto T. Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate. J Biol Chem. 1972 Feb 10;247(3):805–814. [PubMed] [Google Scholar]
  15. Hardy S. J. The stoichiometry of the ribosomal proteins of Escherichia coli. Mol Gen Genet. 1975 Oct 3;140(3):253–274. doi: 10.1007/BF00334270. [DOI] [PubMed] [Google Scholar]
  16. Hirashima A., Wang S., Inouye M. Cell-free synthesis of a specific lipoprotein of the Escherichia coli outer membrane directed by purified messenger RNA. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4149–4153. doi: 10.1073/pnas.71.10.4149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Howard G. A., Traut R. R. Separation and radioautography of microgram quantities of ribosomal proteins by two-dimensional polyacrylamide gel electrophoresis. FEBS Lett. 1973 Jan 15;29(2):177–180. doi: 10.1016/0014-5793(73)80555-1. [DOI] [PubMed] [Google Scholar]
  18. Inouye H., Pratt C., Beckwith J., Torriani A. Alkaline phosphatase synthesis in a cell-free system using DNA and RNA templates. J Mol Biol. 1977 Feb 15;110(1):75–87. doi: 10.1016/s0022-2836(77)80099-5. [DOI] [PubMed] [Google Scholar]
  19. Jaskunas S. R., Lindahl L., Nomura M. Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature. 1975 Oct 9;257(5526):458–462. doi: 10.1038/257458a0. [DOI] [PubMed] [Google Scholar]
  20. Kaltschmidt E., Kahan L., Nomura M. In vitro synthesis of ribosomal proteins directed by Escherichia coli DNA. Proc Natl Acad Sci U S A. 1974 Feb;71(2):446–450. doi: 10.1073/pnas.71.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kaltschmidt E., Wittmann H. G. Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal Biochem. 1970 Aug;36(2):401–412. doi: 10.1016/0003-2697(70)90376-3. [DOI] [PubMed] [Google Scholar]
  22. Kirschbaum J. B., Konrad E. B. Isolation of a specialized lambda transducing bacteriophage carrying the beta subunit gene for Escherichia coli ribonucleic acid polymerase. J Bacteriol. 1973 Nov;116(2):517–526. doi: 10.1128/jb.116.2.517-526.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kung H. F., Fox J. E., Spears C., Brot N., Weissbach H. Studies on the role of ribosomal proteins L 7 and L 12 in the in vitro synthesis of -galactosidase. J Biol Chem. 1973 Jul 25;248(14):5012–5015. [PubMed] [Google Scholar]
  24. Kung H., Spears C., Weissbach H. Purification and properties of a soluble factor required for the deoxyribonucleic acid-directed in vitro synthesis of beta-galactosidase. J Biol Chem. 1975 Feb 25;250(4):1556–1562. [PubMed] [Google Scholar]
  25. LEBOY P. S., COX E. C., FLAKS J. G. THE CHROMOSOMAL SITE SPECIFYING A RIBOSOMAL PROTEIN IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1964 Dec;52:1367–1374. doi: 10.1073/pnas.52.6.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  27. 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]
  28. Lindahl L., Jaskunas S. R., Dennis P. P., Nomura M. Cluster of genes in Escherichia coli for ribosomal proteins, ribosomal RNA, and RNA polymerase subunits. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2743–2747. doi: 10.1073/pnas.72.7.2743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lindahl L., Post L., Nomura M. DNA-dependent in vitro synthesis of fibosomal proteins, protein elongation factors, and RNA polymerase subunit alpha: inhibition by ppGpp. Cell. 1976 Nov;9(3):439–448. doi: 10.1016/0092-8674(76)90089-1. [DOI] [PubMed] [Google Scholar]
  30. Lodish H. F. Translational control of protein synthesis. Annu Rev Biochem. 1976;45:39–72. doi: 10.1146/annurev.bi.45.070176.000351. [DOI] [PubMed] [Google Scholar]
  31. Miller D. L., Weissbach H. An interaction between the transfer factors required for protein synthesis. Arch Biochem Biophys. 1969 Jun;132(1):146–150. doi: 10.1016/0003-9861(69)90346-4. [DOI] [PubMed] [Google Scholar]
  32. Miller D. L., Weissbach H. Studies on the purification and properties of factor Tu from E. coli. Arch Biochem Biophys. 1970 Nov;141(1):26–37. doi: 10.1016/0003-9861(70)90102-5. [DOI] [PubMed] [Google Scholar]
  33. Morrissey J. J., Weissbach H., Brot N. The identification and characterization of proteins similar to L7, L12 in ribosome-free extracts of Escherichia coli. Biochem Biophys Res Commun. 1975 Jul 8;65(1):293–302. doi: 10.1016/s0006-291x(75)80092-1. [DOI] [PubMed] [Google Scholar]
  34. Möller W., Groene A., Terhorst C., Amons R. 50-S ribosomal proteins. Purification and partial characterization of two acidic proteins, A 1 and A 2, isolated from 50-S ribosomes of Escherichia coli. Eur J Biochem. 1972 Jan 31;25(1):5–12. doi: 10.1111/j.1432-1033.1972.tb01660.x. [DOI] [PubMed] [Google Scholar]
  35. Parish J. H., Khairul Bashar S. A., Brown N. L., Brown M. Cell-free synthesis of tryptophanase from Escherichia coli. Use of ribonucleic acid isolated from induced cells and a comparison of the product from a system employing ribosomes with that from one employing ribosomes and exogenous ribonucleic acid. Biochem J. 1971 Nov;125(2):643–653. doi: 10.1042/bj1250643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pettersson I., Hardy S. J., Liljas A. The ribosomal protein L8 is a complex L7/L12 and L10. FEBS Lett. 1976 Apr 15;64(1):135–138. doi: 10.1016/0014-5793(76)80267-0. [DOI] [PubMed] [Google Scholar]
  37. Ramagopal S. Accumulation of free ribosomal proteins S1, L7, and L12 in Escherichia coli. Eur J Biochem. 1976 Oct 1;69(1):289–297. doi: 10.1111/j.1432-1033.1976.tb10885.x. [DOI] [PubMed] [Google Scholar]
  38. Reiness G., Yang H. L., Zubay G., Cashel M. Effects of guanosine tetraphosphate on cell-free synthesis of Escherichia coli ribosomal RNA and other gene products. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2881–2885. doi: 10.1073/pnas.72.8.2881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Reiness G., Zubay G. The messenger-directed synthesis of the alpha-fragment of the enzyme beta-galactosidase. Biochem Biophys Res Commun. 1973 Aug 6;53(3):967–974. doi: 10.1016/0006-291x(73)90186-1. [DOI] [PubMed] [Google Scholar]
  40. Rice R. H., Means G. E. Radioactive labeling of proteins in vitro. J Biol Chem. 1971 Feb 10;246(3):831–832. [PubMed] [Google Scholar]
  41. Salser W., Gesteland R. F., Bolle A. In vitro synthesis of bacteriophage lysozyme. Nature. 1967 Aug 5;215(5101):588–591. doi: 10.1038/215588a0. [DOI] [PubMed] [Google Scholar]
  42. Schumacher G., Ehring R. RNA-directed cell-free synthesis of the galactose enzymes of Escherichia coli. Mol Gen Genet. 1973 Aug 28;124(4):329–344. doi: 10.1007/BF00267662. [DOI] [PubMed] [Google Scholar]
  43. Schweiger M., Gold L. M. DNA-dependent in vitro synthesis of bacteriophage enzymes. Cold Spring Harb Symp Quant Biol. 1969;34:763–766. doi: 10.1101/sqb.1969.034.01.086. [DOI] [PubMed] [Google Scholar]
  44. Subramanian A. R. Copies of proteins L7 and L12 and heterogeneity of the large subunit of Escherichia coli ribosome. J Mol Biol. 1975 Jun 15;95(1):1–8. doi: 10.1016/0022-2836(75)90330-7. [DOI] [PubMed] [Google Scholar]
  45. Ulbrich B., Nierhaus K. H. Pools of ribosomal proteins in Escherichia coli. Studies on the exchange of proteins between pools and ribosomes. Eur J Biochem. 1975 Sep 1;57(1):49–54. doi: 10.1111/j.1432-1033.1975.tb02275.x. [DOI] [PubMed] [Google Scholar]
  46. Wetekam W., Staack K., Ehring R. DNA-dependent in vitro synthesis of enzymes of the galactose operon of Escherichia coli. Mol Gen Genet. 1971;112(1):14–27. doi: 10.1007/BF00266928. [DOI] [PubMed] [Google Scholar]
  47. van Ooyen A. J., Gruber M., Jorgensen P. The mechanism of action of ppGpp on rRNA synthesis in vitro. Cell. 1976 May;8(1):123–128. doi: 10.1016/0092-8674(76)90193-8. [DOI] [PubMed] [Google Scholar]

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