Abstract
When the trp operon of Escherichia coli contains either of two deletions that fuse the initial portion of the leader region to the distal segments of the trpE gene, novel fusion polypeptides are produced. The new polypeptides are synthesized efficiently both in vivo and in vitro, and their synthesis is subject to repression by trp repressor. Fingerprint analyses of tryptic and chymotryptic digests of the new polypeptides show that both contain trpE polypeptide sequences and, despite their different sizes, share the same N-terminal sequence. Our results suggest that synthesis of the new polypeptides is initiated at the AUG-centered ribosome-binding site in the leader region and proceeds in phase to the region coding for the C-terminal end of the trpE polypeptide.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bertrand K., Squires C., Yanofsky C. Transcription termination in vivo in the leader region of the tryptophan operon of Escherichia coli. J Mol Biol. 1976 May 15;103(2):319–337. doi: 10.1016/0022-2836(76)90315-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Bronson M. J., Squires C., Yanofsky C. Nucleotide sequences from tryptophan messenger RNA of Escherichia coli: the sequence corresponding to the amino-terminal region of the first polypeptide specified by the operon. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2335–2339. doi: 10.1073/pnas.70.8.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Files J. G., Weber K., Miller J. H. Translational reinitiation: reinitiation of lac repressor fragments at three internal sites early in the lac i gene of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Mar;71(3):667–670. doi: 10.1073/pnas.71.3.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HELINSKI D. R., YANOFSKY C. Peptide pattern studies on the wild-type A protein of the tryptophan synthetase of Escherichia coli. Biochim Biophys Acta. 1962 Sep 10;63:10–19. doi: 10.1016/0006-3002(62)90333-5. [DOI] [PubMed] [Google Scholar]
- Hershfield V., Boyer H. W., Chow L., Helinski D. R. Characterization of a mini-ColC1 plasmid. J Bacteriol. 1976 Apr;126(1):447–453. doi: 10.1128/jb.126.1.447-453.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackson E. N., Yanofsky C. Thr region between the operator and first structural gene of the tryptophan operon of Escherichia coli may have a regulatory function. J Mol Biol. 1973 May 5;76(1):89–101. doi: 10.1016/0022-2836(73)90082-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Lee F., Yanofsky C. Transcription termination at the trp operon attenuators of Escherichia coli and Salmonella typhimurium: RNA secondary structure and regulation of termination. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4365–4369. doi: 10.1073/pnas.74.10.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Michels C. A., Zipser D. Mapping of polypeptide reinitiation sites within the beta-galactosidase structural gene. J Mol Biol. 1969 May 14;41(3):341–347. doi: 10.1016/0022-2836(69)90280-0. [DOI] [PubMed] [Google Scholar]
- Newton A. Re-initiation of polypeptide synthesis and polarity in the lac operon of Escherichia coli. J Mol Biol. 1969 May 14;41(3):329–339. doi: 10.1016/0022-2836(69)90279-4. [DOI] [PubMed] [Google Scholar]
- Platt T., Squires C., Yanofsky C. Ribosome-protected regions in the leader-trpE sequence of Escherichia coli tryptophan operon messenger RNA. J Mol Biol. 1976 May 15;103(2):411–420. doi: 10.1016/0022-2836(76)90320-x. [DOI] [PubMed] [Google Scholar]
- Platt T., Weber K., Ganem D., Miller J. H. Translational restarts: AUG reinitiation of a lac repressor fragment. Proc Natl Acad Sci U S A. 1972 Apr;69(4):897–901. doi: 10.1073/pnas.69.4.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ray P., Murialdo H. The role of gene Nu3 in bacteriophage lambda head morphogenesis. Virology. 1975 Mar;64(1):247–263. doi: 10.1016/0042-6822(75)90096-3. [DOI] [PubMed] [Google Scholar]
- Sarabhai A., Brenner S. A mutant which reinitiates the polypeptide chain after chain termination. J Mol Biol. 1967 Jul 14;27(1):145–162. doi: 10.1016/0022-2836(67)90357-9. [DOI] [PubMed] [Google Scholar]
- Schmeissner U., Ganem D., Miller J. H. Genetic studies of the lac repressor. II. Fine structure deletion map of the lacI gene, and its correlation with the physical map. J Mol Biol. 1977 Jan 15;109(2):303–326. doi: 10.1016/s0022-2836(77)80036-3. [DOI] [PubMed] [Google Scholar]
- Selker E., Brown K., Yanofsky C. Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColE1. J Bacteriol. 1977 Jan;129(1):388–394. doi: 10.1128/jb.129.1.388-394.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Squires C., Lee F., Bertrand K., Squires C. L., Bronson M. J., Yanofsky C. Nucleotide sequence of the 5' end of tryptophan messenger RNA of Escherichia coli. J Mol Biol. 1976 May 15;103(2):351–381. doi: 10.1016/0022-2836(76)90317-x. [DOI] [PubMed] [Google Scholar]
- Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- Yanofsky C., Horn V., Bonner M., Stasiowski S. Polarity and enzyme functions in mutants of the first three genes of the tryptophan operon of Escherichia coli. Genetics. 1971 Dec;69(4):409–433. doi: 10.1093/genetics/69.4.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zalkin H., Yanofsky C., Squires C. L. Regulated in vitro synthesis of Escherichia coli tryptophan operon messenger ribonucleic acid and enzymes. J Biol Chem. 1974 Jan 25;249(2):465–475. [PubMed] [Google Scholar]
- Zipser D., Zabell S., Rothman J., Grodzicker T., Wenk M. Fine structure of the gradient of polarity in the z gene of the lac operon of Escherichia coli. J Mol Biol. 1970 Apr 14;49(1):251–254. doi: 10.1016/0022-2836(70)90392-x. [DOI] [PubMed] [Google Scholar]