Summary
The DNA dependent synthesis of proteins was studied with a system composed of DNA, washed ribosomes, centrifuged (150000xg) bacterial extract from Escherichia coli and purified initiation factors IF-1 and IF-2. Synthesis of active enzymes encoded by the tryptophan (trp)-operon of E. coli was found to depend strongly on the addition of IF-3, with the same IF-3 dependency for all 5 gene-products of this operon, irrespective of the presence of the promotor proximal gene trpE.
Synthesis of T7 RNA polymerase with T7 DNA as a template, however, was completely independent of the addition of IF-3. The same difference in IF-3 requirement was found when we compared the overall protein synthesis directed by these templates.
This difference could be related to the effect of IF-3 on the formation of initiation complexes with the in vitro prepared mRNA: initiation complexes are readily formed with T7 mRNA also in the absence of IF-3 whereas the formation of these complexes with ϕ80trp mRNA almost completely depends on the presence of this factor.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benne, R., Naaktgeboren, N., Gubbens, J. M., Voorma, H. O.: Recycling of initiation factors IF-1, IF-2, and IF-3. Europ. J. Biochem. 32, 372–380 (1973)
Creighton, T. E., Yanofsky, C.: Indole-3-glycerole-phosphate synthetase of E. coli, an enzyme of the tryptophan operon. J. biol. Chem. 241, 4616–4624 (1966a)
Creighton, T. E., Yanofsky, C.: Association of the α and β2 subunits of the tryptophan synthetase of E. coli. J. biol. Chem. 241, 980–990 (1966b)
Deeb, S. S., Okamoto, K., Hall, B.: Isolation and characterization of non-defective transducing elements of bacteriophage ϕ80. Virology 31, 289–295 (1967)
O'Farrell, P. Z., Gold, I. M.: Transcription and translation of prereplicative bacteriophage T4 genes in vitro. J. biol. Chem. 248, 5512–5519 (1972)
Herrlich, P., Schweiger, M.: RNA polymerase synthesis in vitro directed by Tz phage DNA. Molec. gen. Genet. 110, 31–35 (1971)
Ito, J., Cox, E., Yanofsky, C.: Anthranilate synthetase, an enzyme specified by the tryptophan operon of E. coli. Purification and characterization of component I. J. Bact. 97, 725–733 (1969)
Iwasaki, K., Sabol, S., Wahba, A. J., Ochoa, S.: Translation of the genetic message, VII, role of initiation factors in formation of the chain initiation complex with E. coli ribosomes. Arch. Biochem. Biophys. 125, 542–547 (1968)
Knippenberg, P. H. van: A possible role of the 5' terminal sequence of the 16S ribosomal RNA in the recognition of initiation sequences for protein synthesis. Nucl. Acad. Res. 2, 79–85 (1975)
Leffler, S., Szer, W.: Messenger selection by bacterial ribosomes. Proc. nat. Acad. Sci. (Wash.) 70, 2364–2368 (1973)
Lodish, H. F.: Specificity in bacterial protein synthesis. Role of initiation factors and ribosomal subunits. Nature (Lond.) 226, 705–707 (1970)
Nirenberg, M. W., Leder, P.: RNA codewords and protein synthesis, the effect of trinucleotides upon the binding of sRNA to ribosomes. Science 145, 1399–1407 (1964)
Pouwels, P. H., Rotterdam, J. van: In vitro synthesis of enzymes of the tryptophan operon of E. coli. Proc. nat. Acad. Sci. (Wash.) 69, 1786–1790 (1972)
Pouwels, P. H., Rotterdam, J. van: In vitro synthesis of enzymes of the tryptophan operon of E. coli, studies on the specificity of initiation of transcription. Molec. gen. Genet. 136, 185–197 (1975)
Revel, M., Herzberg, M., Becarevic, A., Gros, F.: Role of a protein factor in the functional binding of ribosomes to natural messenger RNA. J. molec. Biol. 33, 231–249 (1968)
Sabol, S., Sillero, M. A. G., Iwasaki, K., Ochoa, S.: Purification and properties of initiation factor IF-3. Nature (Lond.) 228, 1269–1273 (1970)
Shine, J., Dalgarno, L.: The 3′-terminal sequence of E. coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc. nat. Acad. Sci. (Wash.) 71, 1342–1346 (1974)
Shine, J., Dalgarno, L.: Determinant of cistron specificity in bacterial ribosomes. Nature (Lond.) 254, 34–38 (1975)
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Benne, R., Pouwels, P.H. The role of IF-3 in the translation of T7-and ϕ80trp messenger RNA. Molec. Gen. Genet. 139, 311–319 (1975). https://doi.org/10.1007/BF00267971
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267971