Summary
The nucleotide sequence of the spinach chloroplast 5S rRNA gene and its flanking regions has been determined. A prokaryotic type promoter is to be found upstream of the 5S rRNA gene. Northern blot experiments with selected gene probes show that the 5S gene is co-transcribed with the other ribosomal genes of the operon. This result is confirmed by 5′ S1 mapping of in vivo RNAs synthesised in chloroplasts or in an E. coli strain harboring a mutlicopy plasmid containing the 5S rRNA gene and its flanking regions. In vitro transcription experiments show that initiation of transcription does not occur at the level of the putative 5S rRNA gene promoter. Therefore, we conclude that the 5S rRNA is synthesized only be co-transcription of its gene with the other ribosomal genes of the operon. 3′ S1 nuclease mapping in the spacer region between the 4.5S and the 5S rRNA genes reveals a set of protected fragments located in an A.T rich region downstream of a very stable hairpin and immediately upstream of the putative 5S promoter. This result is interpreted by the presence of preterminated transcripts or processing sites in this region.
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Audren H, Mache R (1986) Nucleic Acids Res 14:9533
Bailey JM, Davidson N (1976) Anal Biochem 70:75–85
Birnboim HC, Doly J (1979) Nucleic Acids Res 7:1513–1523
Bolivar F, Rodrigez RL, Greene RJ, Betlach MC, Heynecker HL, Boyer HW, Crosa JH, Falkow S (1977) Gene 2:95–113
Briat JF, Bisanz-Seyer C, Lescure AM (1987) Curr Genet 11:259–263
Briat JF, Chamberlin MJ (1984) Proc Natl Acad Sci USA 81:7373–7377
Briat JF, Dron M, Loiseaux S, Mache R (1982) Nucleic Acids Res 10:6865–6878
Delihas N, Andersen J, Sprouse HMae, Dudock B (1981) Nucleic Acids Res 9:2801–2805
Dyer TA, Bedbrook JR (1980) Genome organization and expression in plants, vol A29. In: Leaver CJ (ed) Nato Advanced study Institutes series. Plenum, pp 305–312
Edwards K, Bedbrook J, Dyer T, Kossel H (1981) Biochem Int 2:533–538
Gegenheimer P, Apirion D (1981) Microbiol Rev 45:502–541
Gilman MZ, Chamberlin MJ (1983) Cell 35:285–293
Hartley MR (1979) Eur J Biochem 96:311–320
Hawley DK, McClure WR (1983) Nucleic Acids Res 11:2237–2255
Keus RJA, Roovers DJ, Dekker AF, Groot GSP (1983) Nucleic Acids Res 11:3405–3410
Keus RJA, Dekker AF, Kreuk KCJ, Groot GSP (1984) Curr Genet 9:91–97
Kössel H, Edwards K, Koch W, Langridge P, Schiefermayr E, Scharz Zs, Strittmatter G, Zenke G (1982) Nucleic Acids Res (Symp Ser 11):117–120
Lescure AM, Bisanz-Seyer C, Pesey H, Mache R (1985) Nucleic Acids Res 13:8787–8796
Link G, Langridge U (1984) Nucleic Acids Res 12:945–958
Maxam AM, Gilbert W (1977) Proc Natl Acad Sci USA 74:560–564
Messing J (1983) Methods Enzymol 101:143–153
Ohyama K, Fukusawa H, Kohchi T, Shirai H, Sano S, Umesono K, Shiki Y, Takeuchi M, Chang Z, Aota S, Inokuchi H, Oseki H (1986) Nature 322:572–574
Sanger F, Nicklen S, Coulson AR (1977) Proc Natl Acad Sci USA 12:5463–5467
Strittmatter G, Kössel H (1984) Nucleic Acids Res 12:7633–7647
Takaiwa F, Sugiura M (1980) Mol Gen Genet 180:1–4
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Audren, H., Bisanz-Seyer, C., Briat, JF. et al. Structure and transcription of the 5S rRNA gene from spinach chloroplasts. Curr Genet 12, 263–269 (1987). https://doi.org/10.1007/BF00435288
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DOI: https://doi.org/10.1007/BF00435288