Abstract
The enzyme catalyzing the formation of 5-methyluridine (ribothymidine) in tRNA of Escherichia coli is tRNA (uracil-5)-methyltransferase (EC 2.1.1.35). A 2.8-kilobase EcoRI chromosomal DNA fragment contains trmA, the structural gene for this enzyme. Subcloning, transcription in vitro, Tn5 insertion mutagenesis, and transcriptional fusion experiments were performed to establish the gene organization of the trmA region on the E. coli chromosome. trmA is a monocistronic operon. The trmA promoter was localized by in vitro experiments, and the direction of transcription was shown to be counterclockwise on the standard E. coli K-12 chromosomal map. The level of transcription of trmA in vitro and the expression of protein in minicells equal those of the bla gene of plasmid pBR322.
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- Achtman M., Manning P. A., Edelbluth C., Herrlich P. Export without proteolytic processing of inner and outer membrane proteins encoded by F sex factor tra cistrons in Escherichia coli minicells. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4837–4841. doi: 10.1073/pnas.76.10.4837. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Auerswald E. A., Ludwig G., Schaller H. Structural analysis of Tn5. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):107–113. doi: 10.1101/sqb.1981.045.01.019. [DOI] [PubMed] [Google Scholar]
- BERTANI G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol. 1951 Sep;62(3):293–300. doi: 10.1128/jb.62.3.293-300.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Björk G. R. Identification of bacteriophage T4-specific precursor tRNA by using a host mutant defective in the methylation of tRNA. J Virol. 1975 Sep;16(3):741–744. doi: 10.1128/jvi.16.3.741-744.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Björk G. R., Isaksson L. A. Isolation of mutants of Escherichia coli lac king 5-methyluracil in transfer ribonucleic acid or 1-methylguanine in ribosomal RNA. J Mol Biol. 1970 Jul 14;51(1):83–100. doi: 10.1016/0022-2836(70)90272-x. [DOI] [PubMed] [Google Scholar]
- Björk G. R., Olsén A. A method for isolation of Escherichia coli mutants with aberrant RNA methylation using translocatable drug resistance elements. Acta Chem Scand B. 1979;33(8):591–593. doi: 10.3891/acta.chem.scand.33b-0591. [DOI] [PubMed] [Google Scholar]
- Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
- Dürwald H., Hoffmann-Berling H. Endonuclease-I-deficient and ribonuclease I-deficient Escherichia coli mutants. J Mol Biol. 1968 Jul 14;34(2):331–346. doi: 10.1016/0022-2836(68)90257-x. [DOI] [PubMed] [Google Scholar]
- Edlund T., Grundström T., Normark S. Isolation and characterization of DNA repetitions carrying the chromosomal beta-lactamase gene of Escherichia coli K-12. Mol Gen Genet. 1979 Jun 7;173(2):115–125. doi: 10.1007/BF00330301. [DOI] [PubMed] [Google Scholar]
- Gegenheimer P., Apirion D. Processing of procaryotic ribonucleic acid. Microbiol Rev. 1981 Dec;45(4):502–541. doi: 10.1128/mr.45.4.502-541.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greenberg R., Dudock B. Isolation and chracterization of m5U-methyltransferase from Escherichia coli. J Biol Chem. 1980 Sep 10;255(17):8296–8302. [PubMed] [Google Scholar]
- Heller K., Kadner R. J. Nucleotide sequence of the gene for the vitamin B12 receptor protein in the outer membrane of Escherichia coli. J Bacteriol. 1985 Mar;161(3):904–908. doi: 10.1128/jb.161.3.904-908.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hjalmarsson K. J., Byström A. S., Björk G. R. Purification and characterization of transfer RNA (guanine-1)methyltransferase from Escherichia coli. J Biol Chem. 1983 Jan 25;258(2):1343–1351. [PubMed] [Google Scholar]
- Kennedy N., Beutin L., Achtman M., Skurray R., Rahmsdorf U., Herrlich P. Conjugation proteins encoded by the F sex factor. Nature. 1977 Dec 15;270(5638):580–585. doi: 10.1038/270580a0. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Le Grice S. F., Matzura H. Binding of RNA polymerase and the catabolite gene activator protein within the cat promoter in Escherichia coli. J Mol Biol. 1981 Aug 5;150(2):185–196. doi: 10.1016/0022-2836(81)90448-4. [DOI] [PubMed] [Google Scholar]
- Le Grice S. F., Matzura H. Localisation of the transcription initiation site of the chloramphenicol resistance gene on plasmid pAC184. FEBS Lett. 1980 Apr 21;113(1):42–46. doi: 10.1016/0014-5793(80)80490-x. [DOI] [PubMed] [Google Scholar]
- Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
- Marcoli R., Iida S., Bickle T. A. The DNA sequence of an IS/-flanked transposon coding for resistance to chloramphenicol and fusidic acid. FEBS Lett. 1980 Jan 28;110(1):11–14. doi: 10.1016/0014-5793(80)80011-1. [DOI] [PubMed] [Google Scholar]
- Ny T., Björk G. R. Cloning and restriction mapping of the trmA gene coding for transfer ribonucleic acid (5-methyluridine)-methyltransferase in Escherichia coli K-12. J Bacteriol. 1980 May;142(2):371–379. doi: 10.1128/jb.142.2.371-379.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ny T., Björk G. R. Growth rate-dependent regulation of transfer ribonucleic acid (5-methyluridine) methyltransferase in Escherichia coli B/r. J Bacteriol. 1980 Jan;141(1):67–73. doi: 10.1128/jb.141.1.67-73.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ny T., Björk G. R. Stringent regulation of the synthesis of a transfer ribonucleic acid biosynthetic enzyme: transfer ribonucleic acid(m5U)methyltransferase from Escherichia coli. J Bacteriol. 1977 May;130(2):635–641. doi: 10.1128/jb.130.2.635-641.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ny T., Thomale J., Hjalmarsson K., Nass G., Björk G. R. Non-coordinate regulation of enzymes involved in transfer RNA metabolism in Escherichia coli. Biochim Biophys Acta. 1980 Apr 30;607(2):277–284. doi: 10.1016/0005-2787(80)90080-5. [DOI] [PubMed] [Google Scholar]
- Pedersen S., Bloch P. L., Reeh S., Neidhardt F. C. Patterns of protein synthesis in E. coli: a catalog of the amount of 140 individual proteins at different growth rates. Cell. 1978 May;14(1):179–190. doi: 10.1016/0092-8674(78)90312-4. [DOI] [PubMed] [Google Scholar]
- Sakano H., Shimura Y., Ozeki H. Selective modification of nucleosides of tRNA precursors accumulated in a temperature sensitive mutant of Escherichia coli. FEBS Lett. 1974 Nov 1;48(1):117–121. doi: 10.1016/0014-5793(74)81076-8. [DOI] [PubMed] [Google Scholar]
- Schaefer K. P., Altman S., Söll D. Nucleotide modification in vitro of the precursor of transfer RNA of Escherichia coli. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3626–3630. doi: 10.1073/pnas.70.12.3626. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stüber D., Delius H., Bujard H. Electron microscopic analysis of in vitro transcriptional complexes: mapping of promoters of the coliphage T5 genome. Mol Gen Genet. 1978 Oct 30;166(2):141–149. doi: 10.1007/BF00285916. [DOI] [PubMed] [Google Scholar]
- Thompson R., Achtman M. The control region of the F sex factor DNA transfer cistrons: restriction mapping and DNA cloning. Mol Gen Genet. 1978 Oct 24;165(3):295–304. doi: 10.1007/BF00332530. [DOI] [PubMed] [Google Scholar]
- VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]