1887

Abstract

Summary: We have cloned and sequenced a gene of the cyanobacterium sp. strain PCC 6803 named (encoding drug sensory protein A; DspA), mutations in which result in cross-resistance to the herbicides difunon and diuron, as well as to the calmodulin antagonists chlorpromazine and trifluoperazine. The gene encodes a polypeptide of 663 amino acids with a predicted molecular mass of 745 kDa. The molecular nature of two mutations in the gene leading to the cross-resistance has been determined. Targeted mutagenesis of the gene was performed using a kanamycin-resistance gene cartridge. Resulting mutant strains were checked for resistance to difunon and chlorpromazine and showed cross-resistance to both agents. The C-terminal portion of the deduced amino acid sequence of DspA shares significant similarity with the conserved region of histidine protein kinases (HPKs). Hydrophobicity analysis of the amino acid sequence of DspA indicated the existence of two hydrophobic regions in the N-terminal portion that are characteristic of the bacterial sensory HPK family. We suggest that protein DspA is a HPK involved in chemical sensing.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-11-2915
1995-11-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/11/mic-141-11-2915.html?itemId=/content/journal/micro/10.1099/13500872-141-11-2915&mimeType=html&fmt=ahah

References

  1. Aiba H., Nagaya M., Mizuno T. 1993; Sensor and regulator proteins from the cyanobacterium Synechococcus species PCC 7942 that belong to the bacterial signal-transduction families: implication in the adaptive response to phosphate limitation.. Mol Microbiol 8:81–91
    [Google Scholar]
  2. Ajlani G.f, Meyer I., Vernotte C., Astier C. 1989; Mutation in phenol-type herbicide resistance maps within the psbA gene in Synechocystis 6714.. FEBS Lett 246:207–210
    [Google Scholar]
  3. Allen M.M. 1968; Simple conditions for growth of unicellular blue-green algae on plates.. J Phycol 4:1–4
    [Google Scholar]
  4. Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. 1990; Basic local alignment search tool.. Mol Biol 215:403–410
    [Google Scholar]
  5. Chamovitz D., Peker I., Sandmann G., Boger P., Hirschberg J. 1990; Cloning a gene coding for norflurazon resistance in cyanobacteria.. Z Naturforsch Sect C 45:482–486
    [Google Scholar]
  6. Chamovitz D., Peker 1., Hirshberg J. 1991; The molecular basis of resistance to the herbicide norflurazon.. Plant Mol Biol 16:967–974
    [Google Scholar]
  7. Chesnavichene E.A., Elanskaya I.V., Bartsevich V.V., Shestakov S.V. 1994; Molecular analysis of a new gene controlling resistance of the cyanobacterium Synechocystis 6803 to metronidazole and nitrophenolic herbicides.. Dokl Biol Sci 334:18–20
    [Google Scholar]
  8. Chiang G.G., Schaefer M.R., Grossman A.R. 1992; Complementation of a red-light-indifferent cyanobacterial mutant.. Proc Natl Acad Sci USA 89:9415–9419
    [Google Scholar]
  9. Chitnis P.R., Reilly P.A., Miedel M.C., Nelson N. 1989; Structure and targeted mutagenesis of the gene encoding 8-kDa subunit of photosystem I from the cyanobacterium Synechocystis sp. PCC 6803.. J Biol Chem 264:18374–18380
    [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Res 12:387–395
    [Google Scholar]
  11. Engelman D.M., Steitz T.A., Goldman A. 1986; Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins.. Anna Ref Biophys Biophys Chem 15:321–353
    [Google Scholar]
  12. Friedberg D., Seijffers J. 1990; Molecular characterization of genes coding for wild-type and sulfonylurea-resistant acetolactate synthase in the cyanobacterium Synechococcus PCC 7992.. Z Naturforsh Sect C 45:538–543
    [Google Scholar]
  13. Golden S.S., Haselkorn R. 1985; Mutation to herbicide resistance maps within the psbA gene of Anacystis nidulans R2.. Science 229:1104–1107
    [Google Scholar]
  14. Gough J.A., Murray N.E. 1983; Sequence diversity among related genes for recognition of specific targets in DNA molecules.. J Mol Biol 166:1–19
    [Google Scholar]
  15. Grigorieva G., Shestakov S. 1982; Transformation in the cyanobacterium Synechocystis sp. 6803.. FEMS Microbiol Lett 13:367–370
    [Google Scholar]
  16. von Heijne G. 1992; Membrane protein structure prediction, hydrophobicity analysis and the positive-inside rule.. J Mol Biol 225:487–494
    [Google Scholar]
  17. Liang J., Scappino L., Haselkorn R. 1992; The pat A gene product, which contains a region similar to CheY of Escherichia coli, controls heterocyst pattern formation in the cyanobacterium Anabaena 7120.. Proc Natl Acad Sci USA 89:5655–5659
    [Google Scholar]
  18. Mann N.H. 1994; Protein phosphorylation in cyanobacteria.. Microbiology 140:3207–3215
    [Google Scholar]
  19. Martinez-Ferez I.M., Vioque A. 1992; Nucleotide sequence of the phytoene desaturase gene from Synechocystis sp. PCC 6803 and characterization of a new mutation which confers resistance to the herbicide norflurazon.. Plant Mol Biol 18:981–983
    [Google Scholar]
  20. Nagaya M., Aiba H., Mizuno T. 1993; Cloning of a sensory- kinase-encoding gene that belongs to the two-component regu-latory family from the cyanobacterium Synechococcus sp. PCC 7942.. Gene 131:119–124
    [Google Scholar]
  21. Onek L.A., Smith R.J. 1992; Calmodulin and calcium mediated regulation in prokaryotes.. Gen Microbiol 138:1039–1049
    [Google Scholar]
  22. Parkinson J.S., Kofoid E. 1992; Communication modules in bacterial signaling proteins.. Annu Rev Genet 26:71–112
    [Google Scholar]
  23. Reilly P., Hulmes J.D., Pan Y.-C.E., Nelson N. 1988; Molecular cloning and sequencing of the psaD gene encoding subunit II of photosystem I from the cyanobacterium, Synechocystis sp. PCC 6803.. J Biol Chem 263:17658–17662
    [Google Scholar]
  24. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain terminating inhibitors.. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  26. Schluchter W.M., Bryant D.A. 1992; Molecular charac-terization of ferredoxin-NADP+ oxidoreductase in cyanobacteria: cloning and sequence of thepetH gene of Synechococcus sp. PCC 7002 and studies on the gene product.. Biochemistry 31:3092–3102
    [Google Scholar]
  27. Seki T., Yoshikawa H., Takahashi H., Saito H. 1988; Nucleotide sequence of the Bacillus subtilisphoR gene.. J Bacteriol 170:5935–5938
    [Google Scholar]
  28. Shestakov S.V. 1993; Molecular genetic analysis of resistance to the herbicide difunon in the cyanobacterium Synechocystis sp. PCC 6803.. Biol Nauki (in Russian) 2:53–57
    [Google Scholar]
  29. Steinback K.E., Mclntosh L., Bogorad L., Arntzen C.J. 1981; Identification of the triazine receptor protein as a chloroplast gene product.. Proc Natl Acad Sci USA 78:7463–7467
    [Google Scholar]
  30. Stock J.B., Ninfa A.J., Stock A.M. 1989; Protein phosphorylation and regulation of adaptive responses in bacteria.. Microbiol Rev 53:450–490
    [Google Scholar]
  31. Stock J.B., Stock A.M., Mottonen J.M. 1990; Signal transduction in bacteria.. Nature 344:395–400
    [Google Scholar]
  32. Urbach D., Suchanka M., Urbach W. 1976; Effect of substituted pyridazinone herbicides and of difunone (EMD-IT 5914) on carotenoid biosynthesis in green algae.. Z Naturforsch Sect C 31:652–655
    [Google Scholar]
  33. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7- derived system for insertion mutagenesis and sequencing with synthetic universal primers.. Gene 19:259–268
    [Google Scholar]
  34. Williams J.G.K. 1988; Construction of specific mutations in photosystem II photosynthetic reaction center by genetic engineering methods in Synechocystis 6803.. Methods Enzymol 167:766–778
    [Google Scholar]
  35. Yanisch-Perron C, Messing J. 1985; Improved Ml 3 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors.. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-11-2915
Loading
/content/journal/micro/10.1099/13500872-141-11-2915
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error