Abstract
The nucleotide sequence of a 2.3-kilobase-pair DNA fragment of Escherichia coli that contains the transcription signals and the coding region of the pepD gene specifying aminopeptidase D was determined. The location and extent of the open reading frame were verified by partial amino acid sequencing of the purified pepD product. By use of a promoter-screening vector, initiation signals for pepD transcription were located in the 5'-flanking region of the open reading frame. Analysis of pepD transcripts by S1 mapping, primer extension, and Northern (RNA) hybridization revealed two species of monocistronic mRNA with different 5' ends and a common 3' end. Calculation of the degree of codon usage bias in the coding region suggested that the efficiency of pepD translation is relatively low. As deduced from the predicted amino acid sequence, peptidase D is a slightly hydrophilic protein of 485 amino acid residues that contains no extended domains of marked hydrophobicity. Structural and functional features of the pepD gene are discussed and compared with other already sequenced peptidase genes of E. coli.
Full text
PDF










Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bally M., Foglino M., Bruschi M., Murgier M., Lazdunski A. Nucleotide sequence of the promoter and amino-terminal encoding region of the Escherichia coli pepN gene. Eur J Biochem. 1986 Mar 17;155(3):565–569. doi: 10.1111/j.1432-1033.1986.tb09525.x. [DOI] [PubMed] [Google Scholar]
- Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
- Burke J. F. High-sensitivity S1 mapping with single-stranded [32P]DNA probes synthesized from bacteriophage M13mp templates. Gene. 1984 Oct;30(1-3):63–68. doi: 10.1016/0378-1119(84)90105-7. [DOI] [PubMed] [Google Scholar]
- Doolittle R. F., Feng D. F., Johnson M. S., McClure M. A. Relationships of human protein sequences to those of other organisms. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):447–455. doi: 10.1101/sqb.1986.051.01.054. [DOI] [PubMed] [Google Scholar]
- Foglino M., Gharbi S., Lazdunski A. Nucleotide sequence of the pepN gene encoding aminopeptidase N of Escherichia coli. Gene. 1986;49(3):303–309. doi: 10.1016/0378-1119(86)90366-5. [DOI] [PubMed] [Google Scholar]
- Hartmann R. K., Erdmann V. A. Thermus thermophilus 16S rRNA is transcribed from an isolated transcription unit. J Bacteriol. 1989 Jun;171(6):2933–2941. doi: 10.1128/jb.171.6.2933-2941.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
- Henrich B., Schroeder U., Frank R. W., Plapp R. Accurate mapping of the Escherichia coli pepD gene by sequence analysis of its 5' flanking region. Mol Gen Genet. 1989 Feb;215(3):369–373. doi: 10.1007/BF00427031. [DOI] [PubMed] [Google Scholar]
- Ikemura T. Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes: a proposal for a synonymous codon choice that is optimal for the E. coli translational system. J Mol Biol. 1981 Sep 25;151(3):389–409. doi: 10.1016/0022-2836(81)90003-6. [DOI] [PubMed] [Google Scholar]
- Ikemura T., Dahlberg J. E. Small ribonucleic acids of Escherichia coli. I. Characterization by polyacrylamide gel electrophoresis and fingerprint analysis. J Biol Chem. 1973 Jul 25;248(14):5024–5032. [PubMed] [Google Scholar]
- Klein J., Henrich B., Plapp R. Cloning and expression of the pepD gene of Escherichia coli. J Gen Microbiol. 1986 Aug;132(8):2337–2343. doi: 10.1099/00221287-132-8-2337. [DOI] [PubMed] [Google Scholar]
- Konigsberg W., Godson G. N. Evidence for use of rare codons in the dnaG gene and other regulatory genes of Escherichia coli. Proc Natl Acad Sci U S A. 1983 Feb;80(3):687–691. doi: 10.1073/pnas.80.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
- Lazdunski A. M. Peptidases and proteases of Escherichia coli and Salmonella typhimurium. FEMS Microbiol Rev. 1989 Sep;5(3):265–276. doi: 10.1016/0168-6445(89)90035-1. [DOI] [PubMed] [Google Scholar]
- Makino K., Shinagawa H., Amemura M., Nakata A. Nucleotide sequence of the phoB gene, the positive regulatory gene for the phosphate regulon of Escherichia coli K-12. J Mol Biol. 1986 Jul 5;190(1):37–44. doi: 10.1016/0022-2836(86)90073-2. [DOI] [PubMed] [Google Scholar]
- Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller C. G., Schwartz G. Peptidase-deficient mutants of Escherichia coli. J Bacteriol. 1978 Aug;135(2):603–611. doi: 10.1128/jb.135.2.603-611.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murgier M., Gharbi S. Fusion of the lac genes to the promoter for the aminopeptidase N gene of Escherichia coli. Mol Gen Genet. 1982;187(2):316–319. doi: 10.1007/BF00331136. [DOI] [PubMed] [Google Scholar]
- Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
- Nüesch J., Schümperli D. Structural and functional organization of the gpt gene region of Escherichia coli. Gene. 1984 Dec;32(1-2):243–249. doi: 10.1016/0378-1119(84)90052-0. [DOI] [PubMed] [Google Scholar]
- Richardson K. K., Fostel J., Skopek T. R. Nucleotide sequence of the xanthine guanine phosphoribosyl transferase gene of E. coli. Nucleic Acids Res. 1983 Dec 20;11(24):8809–8816. doi: 10.1093/nar/11.24.8809. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
- Sawers G., Böck A. Novel transcriptional control of the pyruvate formate-lyase gene: upstream regulatory sequences and multiple promoters regulate anaerobic expression. J Bacteriol. 1989 May;171(5):2485–2498. doi: 10.1128/jb.171.5.2485-2498.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sharp P. M., Li W. H. Codon usage in regulatory genes in Escherichia coli does not reflect selection for 'rare' codons. Nucleic Acids Res. 1986 Oct 10;14(19):7737–7749. doi: 10.1093/nar/14.19.7737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shinagawa H., Makino K., Nakata A. Regulation of the pho regulon in Escherichia coli K-12. Genetic and physiological regulation of the positive regulatory gene phoB. J Mol Biol. 1983 Aug 15;168(3):477–488. doi: 10.1016/s0022-2836(83)80297-6. [DOI] [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]
- Stirling C. J., Colloms S. D., Collins J. F., Szatmari G., Sherratt D. J. xerB, an Escherichia coli gene required for plasmid ColE1 site-specific recombination, is identical to pepA, encoding aminopeptidase A, a protein with substantial similarity to bovine lens leucine aminopeptidase. EMBO J. 1989 May;8(5):1623–1627. doi: 10.1002/j.1460-2075.1989.tb03547.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stueber D., Ibrahimi I., Cutler D., Dobberstein B., Bujard H. A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences. EMBO J. 1984 Dec 20;3(13):3143–3148. doi: 10.1002/j.1460-2075.1984.tb02271.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace R. B., Miyada C. G. Oligonucleotide probes for the screening of recombinant DNA libraries. Methods Enzymol. 1987;152:432–442. doi: 10.1016/0076-6879(87)52050-x. [DOI] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- Yen C., Green L., Miller C. G. Degradation of intracellular protein in Salmonella typhimurium peptidase mutants. J Mol Biol. 1980 Oct 15;143(1):21–33. doi: 10.1016/0022-2836(80)90122-9. [DOI] [PubMed] [Google Scholar]
- Yoshimoto T., Tone H., Honda T., Osatomi K., Kobayashi R., Tsuru D. Sequencing and high expression of aminopeptidase P gene from Escherichia coli HB101. J Biochem. 1989 Mar;105(3):412–416. doi: 10.1093/oxfordjournals.jbchem.a122678. [DOI] [PubMed] [Google Scholar]