Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of a Regulatory Protein XyIR in the D-Xylose Operon from Escherichia coli

  • Shin, Jae-Ho (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Roh, Dong-Hyun (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Heo, Gun-Young (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Joo, Gil-Jae (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Rhee, In-Koo (Department of Agricultural Chemistry, Kyungpook National University)
  • Published : 2001.12.01
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Abstract

The D-xylose operon in Escherichia coli is known to be regulated by a transcriptional activator protein, XyIR, which is responsible for the expression of both xylAB and xylFGH gene clusters. The XyIR was purified to homogeneity by using the maltose binding protein fusion expression and purification systems involving two chromatography steps. The purified XyIR protein was composed of two subunits of 45 kDa, which was determined by both sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration. The purified XyIR was specifically bounded to the xylA promoter, regardless of adding xylose to the reaction mixture, but binding of XylR was specifically bounded to the xylA promoter, regardless of adding xylose to the reaction mixture, but binding of XylR to the xylA promoter was enhanced by adding xylose. The enhanced binding ability of XyIR in the presence of xylose was not diminished by adding glucose. The presumed XyIR binding site is located between 120 bp to 100 bp upstream the xylA initiation codon.

Keywords

References

  1. Short Protocols in Molecular Biology, 2nd ed. Ausubel, F. M.;R. Brent;R. E. Kingston;D. D. Moore;J. G. Seidman;J. A. Smith;K. Struhl
  2. J. Mol. Biol. v.200 Selection of DNA binding sites by regulatory proteins. Ⅱ. The binding specificity of cyclic AMP receptor protein to recognition sites Berg, O. G.;P. H. von Hippel
  3. Anal. Biochem. v.72 A rapid and sensitive method for the quantitification of microgram quantities of protein utilizing the principle of protein dye binding Bradford, M. M.
  4. J. Mol. Biol. v.209 Determining residue-base interactions between AraC protein and aral DNA Brunelle, A.;R. Schleif
  5. Biochim. Biophys. Acta v.201 Control of xylose metabolism in escherichia coli David, J. D.;H. Wiesmeyer
  6. Necleic Acids Res. v.21 The XylS/AraC family of regulaors Gallegos, M.;M. Carmen;J. L. Ramos
  7. Proc. Natl. Acad. Sci. USA v.82 A dimer of AraC protein contacts three adjacent major groove regions of the ara site Hendrickson, W.;R. Schleif
  8. Gene v.28 Unidirectional digestion with exonulclease Ⅲ creates targeted break points for DNA sequencing Henikoff, S.
  9. Gene v.96 High efficiency transformation of Escherichia coli with plasmids Inoue, H.;H. Nojima;H. Okayama
  10. Agric. Chem. Biotechnol. v.44 Purification and characterization of a thermostable xylose (glucose) isomerase from Streptomyces chilbaensis J-59 Joom G.-J.;J.-H. Shin;G.-Y. Heo;Y.-Y. Kwak;J.-H. Choi;I.-K. Rhee
  11. Escherichia coli and Salmonella typhimurium: cellular and Molecular Biology Dissimilatory pathways for sugars, polyols, and caboxylates Lin, E. S. S.;F. C. Neidhardt(ed.);J. L. Ingraham(ed.);K. B. Low(ed.);B. Magasanik(ed.);M. Schaechter(ed.)H. E. Umbarger(ed.)
  12. Methods Enzymol. v.101 New M13 vectors for cloning Messing, J.
  13. J. Microbiol. Biotechnol. v.7 Overproduction of Escherichia coli D-xylose isomerase using $P_L$ promoter Park, H.-D.;G.-J. Joo;I.-K. Rhee
  14. Bull. Inst. Genet. Eng. Kyungpook Natl. Univ. v.8 Cloning of xylR, regulatory gene for xylose catabolism in Escherichia coli Roh, D-H.;B.-T. Kang;G.-L. Joo;I-K. Rhee
  15. Mol. Gen. Genet. v.194 Cloning and characterization of xyl genes from Escherichia coli Rosenfeld, s. A.;P. E. Stevis;N. W. Y. Ho
  16. Molecular Cloning: A Laboratory Manual. (2nd ed.) Sambrook, J.;E. F. Fritsch;T. Maniatis
  17. J. Baceriol. v.139 Genetics and regulation of D-xylose utilization in Salmonella typhimurium LT2 Shamanna, D. K.;K. E. Sanderson
  18. Nucleic Acids Res. v.22 Analysis of the Escherichia coli genome. Ⅴ. DNA sequence fo the region from 76.0 to 81.5 min Sofia, J. H.;V. Burland;D. L. Daniels;G. Ⅲ Plunkett;F. R. Blattner
  19. J. Bacteriol. v.179 Organization and regulation of the D-xylose operons in escherichia coli K-12: Xy1R acts as a transcriptional activator Song, S.;C. Park
  20. Arch. Microbiol. v.164 Genetics of pentose-phosphate pathway enzymes of Escherichia coli K-12 Sprenger, G. A.
  21. Receptors Channels v.3 Molecular genetics of a receptor protein for D-xylose, encoded by the gene xylF, in Escherichia coli Sumiya, M.;E. O. Davis;L. C. Packman;T. P. McDonald;P. J. F. Henderson
  22. Microniol. Rev. v.57 Structural, functional, and evolutionary relationships among extraceellular solute-binding receptors of bacteria Tam, R.;M. H. Saier, Jr.
  23. J. Mol. Biol. v.196 Psitive regulation of the Escherichia coli L-rhamnose operon is mediated by the products of tandemly repeated regulatory genes Tobin, J. F.;R. Schleif
  24. J. Mol. Biol. v.211 Purification and properties of RhaR, the positive regulatory of the L-rhamnose operon of Escherichia coli Tobin, J. F.;R. Schleif
  25. J. Microbiol. Biotechnol. v.11 Prediction of the secondary structure of the AgfA subunit of Salmonella enteritidis overexpressed as an MBP-fused protein Won, M.;S. Kim;S. Lee;C. Kim;H. Kim;M. Jun;K. B. Song