Summary
Beginning with a synthetic oligonucleotide probe derived from its amino acid sequence, we have identified, cloned and sequenced the hns gene encoding H-NS, an abundant Escherichia coli 15 kDa DNA-binding protein with a possible histone-like function. The amino acid sequence of the protein deduced from the nucleotide sequence is in full agreement with that determined for H-NS. By comparison of the restriction map of the cloned gene and of its neighboring regions with the physical map of E. coli K12 as well as by hybridization of the hns gene with restriction fragments derived from the total chromosome, we have located the hns gene oriented counterclockwise at 6.1 min on the E. coli chromosome, just before an IS30 insertion element.
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Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening of recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Brombach M, Gualerzi CO, Nakamura Y, Pon CL (1986) Molecular cloning and sequence of the Bacillus stearothermophilus translational initiation factor IF2 gene. Mol Gen Genet 205:97–102
Calogero RA, Pon CL, Gualerzi CO (1987) Chemical synthesis and in vivo hyperexpression of a modular gene coding for Escherichia coli translational initiation factor IF1. Mol Gen Genet 208:63–69
Dalrymple B, Caspers P, Arber W (1984) Nucleotide sequence of the prokaryotic mobile genetic element IS30. EMBO J 3:2145–2149
Drlica K, Rouviere-Yaniv J (1987) Histonelike proteins of bacteria. Microbiol Rev 51:301–319
Falconi M, Gualtieri MT, La Teana A, Losso MA, Pon CL (1988) Proteins from the prokaryotic nucleoid. Primary and quaternary structure of the 15 kDa Escherichia coli DNA binding protein H-NS. Mol Microbiol, in press
Gualerzi CO, Losso MA, Lammi M, Friedrich K, Pawlik RT, Canonaco MA, Gianfranceschi G, Pingoud A, Pon CL (1986) Proteins from the prokaryotic nucleoid. Structural and functional characterization of the Escherichia coli DNA-binding proteins NS(HU) and H-NS. In: Gualerzi CO, Pon CL (eds) Bacterial chromatin. Springer, Heidelberg, pp 101–134
Hadley RG, Hu M, Timmons M, Yun K, Deonier RC (1983) A partial restriction map of the proA-purE region of the Escherichia coli K-12 chromosome. Gene 22:281–287
Kano Y, Yoshino S, Wada M, Yokoyama K, Nobuhara M, Imamoto F (1985) Molecular cloning and nucleotide sequence of the HU-1 gene. Mol Gen Genet 201:360–362
Kano Y, Wada M, Nagase T, Imamoto F (1986) Genetic characterization of the gene hupB encoding the HU-1 protein of Escherichia coli. Gene 45:37–44
Kano Y, Osato K, Wada M, Imamoto F (1987) Cloning and sequencing of the HU-2 gene of Escherichia coli. Mol Gen Genet 209:408–410
Kohara Y, Akiyama K, Isono K (1987) The physical map of the whole E. coli chromosome: Application of a new strategy for rapid analysis and sorting of a large genomic library. Cell 50:495–508
Lammi M, Paci M, Pon CL, Losso MA, Miano A, Pawlik RT, Gianfranceschi GL, Gualerzi CO (1984) Proteins from the prokaryotic nucleoid. Biochemical and 1H-NMR studies of three bacterial histone-like proteins. In: Hubscher U, Spadari S (eds) Proteins involved in DNA replication. Plenum, New York and London, pp 467–477
Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353
Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: Complementarity to nonsense triplets and ribosomal binding sites. Proc Natl Acad Sci USA 71:1342–1346
Smith MG (1967) Isolation of high molecular weight DNA from normal and phage-infected Escherichia coli. Methods Enzymol 12:545–550
Timmons MS, Spear K, Deonier RC (1984) Insertion element IS121 is near proA in the chromosomes of Escherichia coli K12 strains. J Bacteriol 160:1175–1177
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Communicated by H. Hennecke
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Pon, C.L., Calogero, R.A. & Gualerzi, C.O. Identification, cloning, nucleotide sequence and chromosomal map location of hns, the structural gene for Escherichia coli DNA-binding protein H-NS. Mol Gen Genet 212, 199–202 (1988). https://doi.org/10.1007/BF00334684
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DOI: https://doi.org/10.1007/BF00334684