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. 1997 Sep;9(9):1673–1682. doi: 10.1105/tpc.9.9.1673

A novel protein with DNA binding activity from tobacco chloroplast nucleoids.

T Nakano 1, S Murakami 1, T Shoji 1, S Yoshida 1, Y Yamada 1, F Sato 1
PMCID: PMC157042  PMID: 9338968

Abstract

A 41-kD DNA binding protein with a basic pl was purified from chloroplast nucleoids in photomixotrophically cultured tobacco cells, and its amino acid sequence was determined. Using this sequence information, its cDNA (CND41) was isolated, and its nucleotide sequence was determined. The predicted amino acid sequence of CND41 has a transit peptide of 120 amino acids and a mature protein of 382 amino acids. A distinctive helix-turn-helix motif in the lysine-rich N-terminal region of the mature protein and an aspartyl protease active site motif were predicted. Expression of a series of truncated CND41 proteins in Escherichia coli indicated that the lysine-rich region is essential for DNA binding and that CND41 nonspecifically binds chloroplast DNA. Protein gel blot analyses showed CND41 mainly in cells and/or tissues containing nonphotosynthesizing, actively growing plastids. In addition, the accumulation of chloroplast transcripts in these cells and/or tissues (e.g., transcripts for QB binding protein of photosystem II [psbA] and large subunit of ribulose bisphosphate carboxylase [rbcL]) was negatively correlated with the accumulation of CND41. Analyses of cultured cells of transgenic tobacco with reduced CND41 levels showed a higher level of expression of chloroplast genes compared with that of the wild type. We discuss the possible function of CND41 as a negative regulator of chloroplast gene expression.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Broyles S. S., Pettijohn D. E. Interaction of the Escherichia coli HU protein with DNA. Evidence for formation of nucleosome-like structures with altered DNA helical pitch. J Mol Biol. 1986 Jan 5;187(1):47–60. doi: 10.1016/0022-2836(86)90405-5. [DOI] [PubMed] [Google Scholar]
  3. Cerutti H., Osman M., Grandoni P., Jagendorf A. T. A homolog of Escherichia coli RecA protein in plastids of higher plants. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8068–8072. doi: 10.1073/pnas.89.17.8068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Churchill M. E., Travers A. A. Protein motifs that recognize structural features of DNA. Trends Biochem Sci. 1991 Mar;16(3):92–97. doi: 10.1016/0968-0004(91)90040-3. [DOI] [PubMed] [Google Scholar]
  5. Dorman C. J., Ni Bhriain N., Higgins C. F. DNA supercoiling and environmental regulation of virulence gene expression in Shigella flexneri. Nature. 1990 Apr 19;344(6268):789–792. doi: 10.1038/344789a0. [DOI] [PubMed] [Google Scholar]
  6. Gruissem W. Chloroplast gene expression: how plants turn their plastids on. Cell. 1989 Jan 27;56(2):161–170. doi: 10.1016/0092-8674(89)90889-1. [DOI] [PubMed] [Google Scholar]
  7. Göransson M., Sondén B., Nilsson P., Dagberg B., Forsman K., Emanuelsson K., Uhlin B. E. Transcriptional silencing and thermoregulation of gene expression in Escherichia coli. Nature. 1990 Apr 12;344(6267):682–685. doi: 10.1038/344682a0. [DOI] [PubMed] [Google Scholar]
  8. Higgins C. F., Hinton J. C., Hulton C. S., Owen-Hughes T., Pavitt G. D., Seirafi A. Protein H1: a role for chromatin structure in the regulation of bacterial gene expression and virulence? Mol Microbiol. 1990 Dec;4(12):2007–2012. doi: 10.1111/j.1365-2958.1990.tb00559.x. [DOI] [PubMed] [Google Scholar]
  9. Iratni R., Baeza L., Andreeva A., Mache R., Lerbs-Mache S. Regulation of rDNA transcription in chloroplasts: promoter exclusion by constitutive repression. Genes Dev. 1994 Dec 1;8(23):2928–2938. doi: 10.1101/gad.8.23.2928. [DOI] [PubMed] [Google Scholar]
  10. Jang J. C., Sheen J. Sugar sensing in higher plants. Plant Cell. 1994 Nov;6(11):1665–1679. doi: 10.1105/tpc.6.11.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Lam E., Hanley-Bowdoin L., Chua N. H. Characterization of a chloroplast sequence-specific DNA binding factor. J Biol Chem. 1988 Jun 15;263(17):8288–8293. [PubMed] [Google Scholar]
  13. Little M. C., Hallick R. B. Chloroplast rpoA, rpoB, and rpoC genes specify at least three components of a chloroplast DNA-dependent RNA polymerase active in tRNA and mRNA transcription. J Biol Chem. 1988 Oct 5;263(28):14302–14307. [PubMed] [Google Scholar]
  14. Mullet J. E. Dynamic regulation of chloroplast transcription. Plant Physiol. 1993 Oct;103(2):309–313. doi: 10.1104/pp.103.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. O'Halloran T. V. Transition metals in control of gene expression. Science. 1993 Aug 6;261(5122):715–725. doi: 10.1126/science.8342038. [DOI] [PubMed] [Google Scholar]
  16. Owen-Hughes T. A., Pavitt G. D., Santos D. S., Sidebotham J. M., Hulton C. S., Hinton J. C., Higgins C. F. The chromatin-associated protein H-NS interacts with curved DNA to influence DNA topology and gene expression. Cell. 1992 Oct 16;71(2):255–265. doi: 10.1016/0092-8674(92)90354-f. [DOI] [PubMed] [Google Scholar]
  17. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reiss T., Link G. Characterization of transcriptionally active DNA-protein complexes from chloroplasts and etioplasts of mustard (Sinapis alba L.). Eur J Biochem. 1985 Apr 15;148(2):207–212. doi: 10.1111/j.1432-1033.1985.tb08826.x. [DOI] [PubMed] [Google Scholar]
  19. Sato F., Shigematsu Y., Yamada Y. Selection of an atrazine-resistant tobacco cell line having a mutant psbA gene. Mol Gen Genet. 1988 Oct;214(2):358–360. doi: 10.1007/BF00337736. [DOI] [PubMed] [Google Scholar]
  20. Schwabe J. W., Rhodes D. Beyond zinc fingers: steroid hormone receptors have a novel structural motif for DNA recognition. Trends Biochem Sci. 1991 Aug;16(8):291–296. doi: 10.1016/0968-0004(91)90121-b. [DOI] [PubMed] [Google Scholar]
  21. Tanaka K., Muramatsu S., Yamada H., Mizuno T. Systematic characterization of curved DNA segments randomly cloned from Escherichia coli and their functional significance. Mol Gen Genet. 1991 May;226(3):367–376. doi: 10.1007/BF00260648. [DOI] [PubMed] [Google Scholar]
  22. Yang J., Schuster G., Stern D. B. CSP41, a sequence-specific chloroplast mRNA binding protein, is an endoribonuclease. Plant Cell. 1996 Aug;8(8):1409–1420. doi: 10.1105/tpc.8.8.1409. [DOI] [PMC free article] [PubMed] [Google Scholar]

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