Abstract
The transcriptionally active chromosome (TAC) of spinach (Spinacia oleracea L.) chloroplasts has been isolated at a high ionic strength, with low mechanical shearing, by glycerol gradient centrifugation. The properties of the TAC differ from those previously reported for the TAC isolated either from Euglena chloroplasts or from spinach using a low-ionic-strength solubilization medium and gel filtration. The high-salt-isolated TAC is homogenous in density but not in size and contains fewer weakly bound proteins than its lowsalt-isolated homologue. In vitro, it promotes elongation of the RNA chains previously initiated in vivo. Transcription is not limited to the ribosomal DNA. The transcriptional pattern is not strongly affected by the high-salt preparation. Ribonuclease pretreatment of the TAC, prior to the in-vitro transcription, leads to a more than tenfold increase of the transcription activity. These properties are discussed in relation to the structure of the spinach chloroplast chromosome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- cpDNA:
-
chloroplast DNA
- rDNA:
-
ribosomal DNA
- RNAse:
-
ribonuclease
- TAC:
-
transcriptionally active chromosome
- tRNA:
-
transfer RNA
References
Arnon, D. (1974) Copper enzyme isolated chloroplast polyphenoloxidase Beta vulgaris. Plant Physiol. 24, 1–15
Blanc, M., Briat, J.F., Laulhère, J.P. (1981) Influence of the ionic environment on the in vitro transcription of the spinach plastid DNA by a selectively bound RNA-polymerase DNA-complex. Biochem. Biophys. Acta 655, 374–382
Briat, J.F., Dron, M., Loiseaux, S., Mache, R. (1982a) Structure and transcription of the spinach chloroplast rDNA leader region. Nucleic Acids Res. 10, 6865–6878
Briat, J.F., Dron, M., Mache, R. (1983) Is transcription of higher plant chloroplast operons regulated by premature termination? FEBS Lett. 163, 1–5
Briat, J.F., Gigot, C., Laulhère, J.P., Mache, R. (1982b) Visualization of a spinach plastid transcriptionally active DNA-protein complex in a highly condensed structure. Plant Physiol. 69, 1205–1211
Briat, J.F., Laulhère, J.P., Mache, R. (1979) Transcription activity of a DNA-protein complex isolated from spinach plastids. Eur. J. Biochem. 98, 285–292
Briat, J.F., Letoffe, S., Mache, R., Rouvière-Yaniv, J. (1984) Similarity between the bacterial histone-like portein HU and a protein from spinach chloroplasts. FEBS Lett. 172, 75–79
Briat, J.F., Mache, R. (1980) Properties and characterization of a spinach chloroplast RNA polymerase isolated from a transcriptionally active DNA-protein complex. Eur. J. Biochem. 111, 503–509
Darnell, J.E. (1982) Variety in the level of gene control in eukariotic cells. Nature 297, 365–371
Greenberg, B.M., Narita, J.O., De-Luca-Flaherty, C., Gruissem, W., Rushlow, K.A., Hallick, R.B. (1984) Evidence for two RNA polymerase activities in Euglena gracilis chloroplasts. J. Biol. Chem. 259, 14880–14887
Gruissem, W., Greenberg, B.M., Zurawski, G., Prescott, D.M., Hallick, R.B. (1983) Biosynthesis of chloroplast transfer RNA in a spinach chloroplast transcription system. Cell 35, 815–828
Hallick, R.B., Lipper, C., Richards, O.C., Rutter, W.J. (1976) Isolation of a transcriptionally active chromosome from chloroplasts of Euglena gracilis. Biochemistry 15, 3039–3045
Hansmann, P., Falk, H., Ronaì, K., Sitte, P. (1985) Structure, composition and distribution of plastid nucleoids in Narcissus pseudonarcissus. Planta 164, 459–472
Herrmann, R.G., Possingham, J.V. (1980) Plastid DNA, the plastome, in chloroplasts. In: Results and problems in cell differentiation, vol. 10, pp. 45–96, Reiner, J., ed. Springer Verlag, Berlin Heidelberg New York
Jackson, D.A., Cook, P.R. (1985) A general method for preparing chromatin containing intact DNA. — Transcription occurs at a nucleoskeleton. EMBO J. 4, 913–925
Kolodner, R., Warner, R.C., Tewari, K.K. (1975) The presence of covalently linked ribonucleotides in the closed circular deoxyribonucleic acid from higher plants. J. Biol. Chem. 250, 7020–7026
Lerbs, S., Briat, J.F., Mache, R. (1983) Chloroplast RNA polymerase from spinach: purification and DNA-binding proteins. Plant Mol. Biol. 2, 67–74
Maaloe, O., Kjeldgaard, N. (1966) Control of macromolecular synthesis, pp. 188–197. W.A. Benjamin, New York
Maniatis, T., Fritsche, F., Sambrook, J. (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Narita, J.O., Rushlow, K.E., Hallick, R.B. (1985) Characterization of a Euglena gracilis chloroplast RNA polymerase specific for ribosomal RNA genes. J. Biol. Chem. 260, 11194–11199
Pettijohn, D.E., Clarkson, K., Kossman, C.R., Stonington, O.G. (1970) Synthesis of ribosomal RNA on a protein-DNA complex isolated from bacteria: a comparison of ribosomal RNA synthesis in vitro. J. Mol. Biol. 52, 281–300
Reiss, T., Link, G. (1985) Characterization of transcriptionally active DNA-protein complex from chloroplasts of mustard (Sinapis alba L.). Eur. J. Biochem. 148, 207–212
Rushlow, K.E., Orozco, E.M., Lipper, C., Hallick, R.B. (1980) Selective in vitro transcription of Euglena chloroplast ribosomal RNA genes by a transcriptionally active chromosome. J. Biol. Chem. 255, 3786–3792
Sacchi, A., Ferrini, U., Londei, P., Cammarano, P., Maraldi, N. (1977) Mitochondrial and cytoplasmic ribosomes from mammalian tissues. Further characterization of ribosomal subunits and validity of buoyant density methods for determination of the chemical composition and partial specific volume of ribonucleoprotein particles. Biochem. J. 166, 245–259
Saitoh, T., Ishihama, A. (1977) Biosynthesis of RNA polymerase in Escherichia coli. VI. Distribution of RNA polymerase subunits between nucleoid and cytoplasm. J. Mol. Biol. 115, 403–416
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lebrun, M., Briat, J.F. & Laulhere, J.P. Characterization and properties of the spinach chloroplast transcriptionally active chromosome isolated at high ionic strength. Planta 169, 505–512 (1986). https://doi.org/10.1007/BF00392099
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392099