Abstract
Approximately two-thirds of the 55 to 60 plastid ribosomal proteins are encoded in the nucleus. Since the protein products of each of these genes are needed in equal amounts for ribosome assembly, their expression may be coordinately regulated by common mechanisms. To begin to understand how the expression of these genes is regulated, we have isolated cDNA and genomic clones for three plastid ribosomal protein genes from an Arabidopsis thaliana library. The genes rps17, rpl9 and rpl15, encoding plastid ribosomal proteins CS17, CL9 and CL15, respectively, are located in the nuclear genome and Southern blot data suggest that each is a single copy gene in A. thaliana. Northern blot data show that transcripts from rps17, rpl9 and rpl15 are much more abundant in leaves and stems than they are in roots. The nucleotide sequences of each of these three genes were determined and their transcriptional initiation sites identified. rps17 transcripts have multiple 5′ ends suggesting that they are initiated at multiple sites or are post-transcriptionally processed at their 5′ end. rpl9 and rpl15 apparently have unique transcriptional initiation sites but are post-transcriptionally processed to remove six and three introns, respectively, from their primary transcripts. We have examined the genomic sequences for motifs that may be important for the proper expression of these genes. A 7 bp sequence motif, whose consensus is 5′-AGGCCCA-3′, flanked by AT-rich regions was identified between 38 and 73 nucleotides upstream of the rps17, rpl9 and rpl15 transcriptional initiation sites.
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References
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: Current Protocols in Molecular Biology, volume 1. Greene Publishing Associates/Wiley-Interscience (1989).
Baulcombe DC, Key JL: Polyadenylated RNA sequences which are reduced in concentration following auxin treatment of soybean hypocotyls. J. Biol Chem 255: 8907–8913 (1980).
Bendich AJ, Anderson RS, Ward BL: Plant DNA: Long, pure and simple. In: Leaver CJ (ed) Genome Organization and Expression in Plants, pp. 31–33. Plenum Press, New York (1980).
Bisanz-Seyer C, Li Y, Seyer P, Mache R: The components of the plastid ribosome are not accumulated synchronously during the early development of spinach plants. Plant Mol Biol 12: 201–211 (1989).
Bonham-Smith PC, Bourque DP: The chloroplast genome and regulation of its expression. In: Adolph KW (ed) Chromosomes: Eukaryotic, Prokaryotic, and Viral, Vol 2, pp. 180–216. CRC Press, Boca Raton, FL (1990).
Boynton JE, Gillham NW, Lambowitz AM: Biogenesis of chloroplast and mitochondrial ribosomes. In: Chambliss G, Craven GR, Davies J, Davis K, Kahan L, Nomura M (eds), Ribosomes: Structure, Function and Genetics, pp. 903–950. University Park Press, Baltimore (1979).
Calzone FJ, Britten RJ, Davidson EH: Mapping of gene transcripts by nuclease protection assays and cDNA primer extension. Meth Enzymol 152: 611–632 (1987).
Dynan WS: Promoters for housekeeping genes. Trends Genet 2: 196–197 (1986).
Elliston K, Messing J: The molecular architecture of plant genes: A phylogenetic perspective. In: Kahl G (ed) Architecture of Eukaryotic Genes, pp. 229–240. VCH Verlagsgesellschaft, New York (1988).
Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).
Firestein GS, Gardner SM, Roeder WD: Quantitative molecular hybridization with unfractionated solubilized cells using RNA probes and polyacrylamide gel electrophoresis. Anal Biochem 167: 381–386 (1987).
Gantt JS, Key JL: Isolation of nuclear-encoded plastid ribosomal protein cDNAs. Mol Gen Genet 202: 186–193 (1986).
Gantt JS: Nucleotide sequences of cDNAs encoding four complete nuclear-encoded plastid ribosomal proteins. Curr Genet 14: 519–528 (1988).
Gantt JS, Thompson MD: Plant cytosolic ribosomal protein S11 and chloroplast ribosomal protein CS17: Their primary structures and evolutionary relationships. J. Biol Chem 265: 2763–2767 (1990).
Gantt JS, Gupta A, Thompson MD: The effects of chlorophyll photooxidation on nuclear-encoded plastid ribosomal protein mRNAs in Norflurazon-treated pea seedlings. In: Mache R, Stutz E, Subramanian AR (eds) The Translational Apparatus of Photosynthetic Organelles, pp. 207–213. Springer-Verlag, Berlin (1991).
Gigot C: Histone genes in higher plants. In: Kahl G (ed), Architecture of Eukaryotic Genes, pp. 229–240. VCH Verlagsgesellschaft, New York (1988).
Giorginis S, Chen R: The primary structure of protein L15 located at the peptidyl-transferase center of Escherichia coli ribosomes. FEBS Lett 84: 347–350 (1977).
Gorenstein C, Warner JR: Coordinate regulation of the synthesis of eukaryotic ribosomal proteins. Proc Natl Acad Sci USA 73: 1547–1551 (1976).
Hanley BA, Schuler MA: Plant intron sequences: Evidence for distinct groups of introns. Nucl Acids Res 16: 7159–7176 (1988).
Hariharan N, Kelley DE, Perry RP: Equipotent mouse ribosomal protein promoters have a similar architecture that includes internal sequence elements. Genes Devel 3: 1789–1800 (1989).
Hatakeyama T, Kaufmann F, Schroeter B: Primary structures of five ribosomal proteins from the archaebacterium Halobacterium marismortui and their structural relationships to eubacterial and eukaryotic ribosomal proteins. Eur J Biochem 185: 685–693 (1989).
Henikoff S: Unidirectional digestion with exonuclease III in DNA sequence analysis. Meth Enzymol 155: 156–165 (1987).
Johnson CH, Subramanian AR: Chloroplast ribosomal protein L15, like L1, L13 and L21, is significantly larger than its E. coli homologue. FEBS Lett 282: 268–272 (1991).
Joshi CP: An inspection of the domain between putative TATA box and translation start sites in 79 plant genes. Nucl Acids Res 15: 6643–6653 (1987).
Kamp RM, Wittmann-Liebold B: The primary structure of protein L9 from the Escherichia coli ribosome. FEBS Lett 149: 313–319 (1982).
Käufer NF, Fried HM, Schwindinger WF, Jasin M, Warner JR: Cycloheximide resistance in yeast: The gene and its protein. Nucl Acids Res 11: 3123–3135 (1983).
Keegstra K, Olsen LJ, Theg SM: Chloroplastic precursors and their transport across the envelope membranes. Annu Rev Plant Physiol Plant Mol Biol 40: 471–501 (1989).
Kim Y, Zhang H, Scholl RL: Two evolutionarily divergent genes encode a cytoplasmic ribosomal protein of Arabidopsis thaliana. Gene 93: 177–182 (1990).
Larkin CJ, Hunsperger JP, Culley D, Rubenstein I, Silflow CD: The organization and expression of a maize ribosomal protein gene family. Genes Devel 3: 500–509 (1989).
Leer RJ, vanRaamsdonk-Duin MMC, Mager WH, Planta RJ: Conserved sequences upstream of yeast ribosomal protein genes. Curr Genet 9: 273–277 (1985).
Lotti M, Dabbs ER, Hasenbank R, Stöffler-Meilicke M, Stöffler G: Characterisation of a mutant from Escherichia coli lacking protein L15 and localisation of protein L15 by immuno-electron microscopy. Mol Gen Genet 192: 295–300 (1983).
Mache R: Chloroplast ribosomal proteins and their genes. Plant Sci 72: 1–12 (1990).
Madsen LH, Kreiberg JD, Gausing K: A small gene family in barley encodes ribosomal proteins homologous to yeast YL17 and L22 from archaebacteria, eubacteria, and chloroplast. Curr Genet 19: 417–422 (1991).
Mager WH: Control of ribosomal protein expression. Biochim Biophys Acta 949: 1–15 (1988).
Mager WH, Planta RJ: Multifunctional DNA-binding proteins mediate concerted transcription activation of yeast ribosomal protein genes. Biochim Biophys Acta 1050: 351–355 (1990).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Margulis L, Bermudes D: Symbiosis as a mechanism of evolution: status of cell symbiosis theory. Symbiosis 1: 101–124 (1985).
Mermelstein FH, Flores O, Reinberg D: Initiation of transcription by RNA polymerase II. Biochim Biophys Acta 1009: 1–10 (1989).
Mullet JE: Chloroplast development and gene expression. Annu Rev Plant Physiol Plant Mol Biol 39: 475–502 (1988).
Nomura M, Gourse R, Baughman G: Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem 53: 75–117 (1984).
Olszewski NE, Martin FB, Ausubel FM: Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene Nicotiana tabacum. Nucl Acids Res 16: 10765–10782 (1988).
Palmer JD: Comparative organization of chloroplast genomes. Annu Rev Genet 19: 823–833 (1985).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Schnier J, Kitakawa M, Isono K: The nucleotide sequence of an E. coli chromosomal region containing the genes for ribosomal proteins S6, S18, L9 and an open reading frame. Mol Gen Genet 204: 126–132 (1986).
Wang Y: Double-stranded DNA sequencing with T7 polymerase. Biotechniques 6: 843–845 (1988).
Warner JR: Synthesis of ribosomes in Saccharomyces cerevisiae. Microbiol Rev 53: 256–271 (1989).
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Thompson, M.D., Jacks, C.M., Lenvik, T.R. et al. Characterization of rps17, rpl9 and rpl15: three nucleus-encoded plastid ribosomal protein genes. Plant Mol Biol 18, 931–944 (1992). https://doi.org/10.1007/BF00019207
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DOI: https://doi.org/10.1007/BF00019207