Abstrct
Plastids, the characteristic organelles of plant cells, contain their own DNA and a complete apparatus for transcription and translation (1). Although this is principally similar to what is known for mitochondria, the details of the plastid gene expression system appear to be more complex, both with regard to the number of genes and mechanisms involved. Moreover, unlike mitochondria, plastids exist in various tissue-specific and developmental stage-specific forms, each differing in its state of gene expression. The best-known plastid types are the chloroplasts found in green (lightgrown) tissue and the etioplasts that develop in higher plant seedlings when these are grown in the dark (2). In contrast to the mitochondrial RNA polymerase enzyme, highly purified chloroplast enzyme preparations consist of a multisubunit complex similar to those in bacteria and also in the nuclei of eukaryotic cells, but with considerable species-specific variations. It is still an open question whether multiple entirely distinct RNA polymerases exist within the chloroplast and other plastid forms. Alternatively, one common core enzyme could be surrounded by different specificity factors, which would account for multiple, but closely related, holoenzymes.
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References
Bogorad, L. and Vasil, I. K. (1991) The Molecular Biology of Plastids. Academic, New York.
Link, G. (1991) Photoregulated development of chloroplasts, in The Photosynthetic Apparatus: Molecular Biology and Operation (Bogorad, L. and Vasil, I. K., eds.), Academic, New York, pp. 365–394.
Hallick, R., Lipper, C., Richards, O. C., and Rutter, W. J. (1976) Isolation of a transcriptionally active chromosome from chloroplasts of Euglena gracilis. Bio-chemistry 15, 3039–3045.
Briat, J. F., Laulhere, J. P., and Mache, R. (1979) Transcription activity of a DNA-protein complex isolated from spinach plastids. Eur. J. Biochem. 98, 285–292.
Reiss, T. and Link, G. (1985) Characterization of transcriptionally active DNA-protein complexes from chloroplasts and etioplasts of mustard (Sinapis alba L.). Eur. J. Biochem. 148, 207–212.
Hu, J. and Bogorad, L (1990) Maize chloroplast RNA polymerase: the 180-and 38-kilodalton polypeptides are encoded in chloroplast genes. Proc. Natl. Acad. Sci. USA 87, 1531–1535.
Hu, J., Troxler, R. F., and Bogorad, L. (1991) Maize chloroplast RNA polymerase the 78-kilodalton polypeptide is encoded by the plastid rpoCl gene. Nucleic Acids Res. 19, 3431–3434.
Eisermann, A., Tiller, K., and Link, G. (1990) In vitro transcription and DNA binding characteristics of chloroplast and etioplast extracts from mustard (Sinapis alba) indicate differential usage of the psbA promoter. EMBO J. 9, 3981–3987.
Bülow, S. and Link, G. (1988) Sigma-like activity from mustard (Sinapis alba L.) chloroplasts conferring DNA-binding and transcription specificity to E. coli core RNA polymerase. Plant Mol. Biol. 10, 349–357.
Tiller, K., Eisermann, A., and Link, G.(1991) The chloroplast transcription apparatus from mustard (Sinapis alba L.). Evidence for three different transcription factors which resemble bacterial sigma factors. Eur. J. Biochem. 198, 93–99.
Bottomley, W., Smith, H. J., and Bogorad, L. (1971) RNA polymerases of maize: partial purification and properties of the chloroplast enzyme. Proc. Natl. Acad. Sci. USA 68, 2412–2416.
Bickle, T. A., Pirrotta, V., and Imber, R. (1977) A simple general procedure for purifying restriction endonucleases. Nucleic Acids Res. 4, 2561–2572.
Briat, J. F., Dron, M., Loiseaux, S., and Mache, R. (1982) Structure and transcription of the spinach chloroplast rDNA leader region. Nucleic Acids Res. 10, 6865–6877.
Lerbs, S., Briat, J. F., and Mache, R. (1983) Chloroplast RNA polymerase from spinach: purification and DNA-binding proteins. Plant Mol. Biol. 2, 67–74.
Sambrook, J., Fritsch, E. F., and Maniatis, T.(1989) Molecular Cloning, A Luboratory Manual, 2nd ed, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Billow, S., Reiss, T., and Link, G. (1987) DNA-binding proteins of the transcriptionally active chromosome from mustard (Sinapis alba L.) chloroplasts. Curr. Genet. 12, 157–159.
Khanna, N. C., Lakhani, S., and Tewari, K.K., (1992) Identification of the template binding polypeptide in the pea chloroplast transcriptional complex. Nucleic Acids Res. 20, 69–74.
Lam, E., Hanley-Bowdoin, L., and Chua, N. H. (1988) Characterization of a chloroplast sequence-specific DNA binding factor. J Biol. Chem 263, 8288–8293.
Zaitlin, D., Hu, J., and Bogorad, L. (1989) Binding and transcription of relaxed DNA templates by fractions of maize chloroplast extracts. Proc. Natl. Acad. Sci. USA 86, 876–880.
Fried, M. and Crothers, D. M. (1981) Equilibria and kinetics of lac repressor-operator interaction by polyacrylamide gel electrophoresis. Nucleic Acids Res. 9, 6505–6523.
Garner, M. M. and Revzin, A. (1981) A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 9, 3047–3060.
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© 1995 Humana Press Inc.
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Tiller, K., Link, G. (1995). Plastid In Vitro Transcription. In: Tymms, M.J. (eds) In Vitro Transcription and Translation Protocols. Methods in Molecular Biology, vol 37. Humana Press. https://doi.org/10.1385/0-89603-288-4:121
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DOI: https://doi.org/10.1385/0-89603-288-4:121
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