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J Bacteriol. Aug 1988; 170(8): 3593–3600.
PMCID: PMC211333

Cloning and characterization of Bacillus subtilis iep, which has positive and negative effects on production of extracellular proteases.


We have isolated a DNA fragment from Bacillus subtilis 168 which, when present in a high-copy plasmid, inhibited production of extracellular alkaline and neutral proteases. The gene responsible for this activity was referred to as iep. The open reading frame of iep was found to be incomplete in the cloned DNA fragment. When the intact iep gene was reconstructed after the missing part of the iep gene had been cloned, it showed an enhancing effect on the production of the extracellular proteases. The open reading frame encodes a polypeptide of 229 amino acids with a molecular weight of ca. 25,866. Deletion of two amino acids from the N-terminal half of the putative iep protein resulted in dual effects, i.e., a decrease in the inhibitory activity shown by the incomplete iep gene and a slight increase in the enhancing activity shown by the complete iep gene. These results show that the iep gene product is a bifunctional protein, containing inhibitory and enhancing activities for the exoprotease production in the N-terminal and C-terminal regions, respectively. It was found by genetic and functional analyses that iep lies very close to sacU.

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Selected References

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  • Amory A, Kunst F, Aubert E, Klier A, Rapoport G. Characterization of the sacQ genes from Bacillus licheniformis and Bacillus subtilis. J Bacteriol. 1987 Jan;169(1):324–333. [PMC free article] [PubMed]
  • Chang S, Cohen SN. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. [PubMed]
  • Gaur NK, Dubnau E, Smith I. Characterization of a cloned Bacillus subtilis gene that inhibits sporulation in multiple copies. J Bacteriol. 1986 Nov;168(2):860–869. [PMC free article] [PubMed]
  • Hahn J, Albano M, Dubnau D. Isolation and characterization of Tn917lac-generated competence mutants of Bacillus subtilis. J Bacteriol. 1987 Jul;169(7):3104–3109. [PMC free article] [PubMed]
  • Hattori M, Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Higerd TB, Hoch JA, Spizizen J. Hyperprotease-producing mutants of Bacillus subtilis. J Bacteriol. 1972 Nov;112(2):1026–1028. [PMC free article] [PubMed]
  • Hoch SO, Crawford IP. Enzymes of the tryptophan pathway in three Bacillus species. J Bacteriol. 1973 Nov;116(2):685–693. [PMC free article] [PubMed]
  • Horinouchi S, Weisblum B. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982 May;150(2):815–825. [PMC free article] [PubMed]
  • Kawamura F, Doi RH. Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. J Bacteriol. 1984 Oct;160(1):442–444. [PMC free article] [PubMed]
  • Lederberg EM, Cohen SN. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. [PMC free article] [PubMed]
  • Lepesant JA, Kunst F, Lepesant-Kejzlarová J, Dedonder R. Chromosomal location of mutations affecting sucrose metabolism in Bacillus subtilis Marburg. Mol Gen Genet. 1972;118(2):135–160. [PubMed]
  • Nagami Y, Tanaka T. Molecular cloning and nucleotide sequence of a DNA fragment from Bacillus natto that enhances production of extracellular proteases and levansucrase in Bacillus subtilis. J Bacteriol. 1986 Apr;166(1):20–28. [PMC free article] [PubMed]
  • Pabo CO, Sauer RT. Protein-DNA recognition. Annu Rev Biochem. 1984;53:293–321. [PubMed]
  • Piggot PJ, Hoch JA. Revised genetic linkage map of Bacillus subtilis. Microbiol Rev. 1985 Jun;49(2):158–179. [PMC free article] [PubMed]
  • Rosenberg M, Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. [PubMed]
  • Sadaie Y, Kada T. Bacillus subtilis gene involved in cell division, sporulation, and exoenzyme secretion. J Bacteriol. 1985 Aug;163(2):648–653. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Southern E. Gel electrophoresis of restriction fragments. Methods Enzymol. 1979;68:152–176. [PubMed]
  • Steinmetz M, Kunst F, Dedonder R. Mapping of mutations affecting synthesis of exocellular enzymes in Bacillus subtilis. Identity of the sacUh, amyB and pap mutations. Mol Gen Genet. 1976 Nov 17;148(3):281–285. [PubMed]
  • Sutcliffe JG. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. [PubMed]
  • Tanaka T, Kawano N. Cloning vehicles for the homologous Bacillus subtilis host-vector system. Gene. 1980 Jul;10(2):131–136. [PubMed]
  • Ward JB, Jr, Zahler SA. Genetic studies of leucine biosynthesis in Bacillus subtilis. J Bacteriol. 1973 Nov;116(2):719–726. [PMC free article] [PubMed]
  • Yang M, Ferrari E, Chen E, Henner DJ. Identification of the pleiotropic sacQ gene of Bacillus subtilis. J Bacteriol. 1986 Apr;166(1):113–119. [PMC free article] [PubMed]
  • Yoneda Y, Maruo B. Mutation of Bacillus subtilis causing hyperproduction of alpha-amylase and protease, and its synergistic effect. J Bacteriol. 1975 Oct;124(1):48–54. [PMC free article] [PubMed]
  • Zuber P, Losick R. Role of AbrB in Spo0A- and Spo0B-dependent utilization of a sporulation promoter in Bacillus subtilis. J Bacteriol. 1987 May;169(5):2223–2230. [PMC free article] [PubMed]

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