Abstract
The sequence containing `upstream Sal repeats' (USR) from the Arabidopsis thaliana ribosomal DNA intergenic region (IGR) was tested for its influence on the in vivo activity of nearby protein coding genes. On average, the presence of the IGR fragment leads to a four-fold increase in the expression of a reporter gene, β-glucuronidase, under control of the strong CaMV 35S promoter. With the help of the site-specific cre-lox recombination system, we have also obtained pairs of transgenic lines with or without the USR-containing fragment, both integrated at the same chromosomal position. Results with these transgenic lines, which contain an NPT II (kanamycin resistance) gene under control of the nos promoter as a test gene, confirmed the results obtained with the CaMV 35S-driven GUS gene. Moreover, they show that the IGR sequence can oppose tendencies of gene silencing. We hypothesize that the described effect relates to features of the chromatin structure in the proximity of the upstream Sal repeats.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Allen, G.C., Hall, G. Jr., Michalowski, S., Newman, W., Spiker, S., Weissinger, A.K. and Thompson, W.F. 1996. High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco. Plant Cell 8: 899–913.
Ausubel, F.M., Brent, R., Kingston, R.F., Moore, D.O., Seidman, J.G., Smith, J.A. and Struhl, K. 1987. Current Protocols in Molecular Biology. John Wiley, New York.
Bechthold, N., Ellis, J. and Pelletier, G. 1993. In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C.R. Acad. Sci. Paris, Life Sci. 316: 1194–1199.
Becker, D. 1990. Binary vectors which allow the exchange of plant selectable markers and reporter genes. Nucl. Acids Res. 18: 203.
Borisjuk, N., Borisjuk, L., Komarnytski, S., Timeva, S., Hemleben, V., Gleba, Y. and Raskin, I. 2000. Tobacco ribosomal DNA spacer element stimulates amplification and expression of heterologous genes. Nature Biotechnol. 18: 1303–1306.
Breyne, P., Gheysen, G., Jacobs, A., Van Montagu, M. and Depicker, A. 1992. Effect of T-DNA configuration on transgene expression. Mol. Gen. Genet. 235: 389–396.
Carmo-Fonseca, M., Mendes-Soares, L. and Campos, I. 2000. To be or not to be in the nucleolus. Nature Cell Biol. 2: E107–E112.
Copenhaver, G.P., Doelling, J.H., Gens, J.S. and Pikaard, C.S. 1995. Use of RFLPs larger than 100 kbp to map the position and internal organization of the nucleolus organizer region on chromosome 2 in Arabidopsis thaliana. Plant J. 7: 273–286.
Davis, E.S., Shafer, B.K. and Strathern, J.N. 2000. The Saccharomyces cerevisiae RDN1 locus is sequestered from interchromosomal meiotic ectopic recombination in a SIR2-dependent manner. Genetics 155: 1019–1032.
Doelling, J.H. and Pikaard, C.S. 1995. The minimal ribosomal RNA gene promoter of Arabidopsis thaliana includes a critical element at the transcription initiation site. Plant J. 8: 683–692.
Doelling, J.H., Gaudino, R.J. and Pikaard, C.S. 1993. Functional analysis of Arabidopsis thaliana rRNA gene spacer promoters in vivo and by transient expression. Proc. Natl. Acad. Sci. USA 90: 7528–7532.
Ghosh, A.K., Kermekchiev, M. and Jacob, S.T. 1994. Effects of repetitive and non-repetitive rat rDNA enhancer elements on in vivo transcription by RNA polymerases I and II. Gene 141: 271–275.
González-Melendi, P, Wells, B., Beven, A.F. and Shaw, P.J. 2001. Single ribosomal transcription units are linear, compacted christmas trees in plant nucleoli. Plant J. 27: 223–233.
Gründler, P., Unfried, I., Pascher, K. and Schweizer, D. 1991. rDNA intergenic region from Arabidopsis thaliana: structural analysis, interspecific variation and functional implications. J. Mol. Biol. 221: 1209–1222.
Gründler, P., Unfried, I., Pointner, R. and Schweizer, D. 1989Nucleotide sequence of the 25S-18S ribosomal gene spacer from Arabidopsis thaliana. Nucl. Acids Res. 17: 6395–6396.
Hemleben, V. and Zentgraf, U. 1994. Structural organization and regulation of transcription by RNA polymerase I of plant nuclear ribosomal RNA genes. In: L. Nover (Ed.) Results and Problems of Cell Differentiation, Vol. 20, Springer-Verlag, Berlin. pp. 3–24.
Jacob, S.T. and Ghosh, A.K. 1999. Control of RNA polymerase I-directed transcription: recent trends. J. Cell. Biochem. Suppl. 32/33: 41–50.
Kobayashi, T., Nomura, M. and Horiuchi, T. 2001. Identification of DNA cis elements essential for expansion of ribosomal DNA repeats in Saccharomyces cerevisiae. Mol. Cell. Biol. 21: 136–147.
Längst, G., Becker, P.B. and Grummt, I. 1994. TTF-I determines the chromatin architecture of the active rDNA promoter. EMBO J. 17: 3135–3145.
Martin, T., Wöhner, R.-V., Hummel, S., Willmitzer, L. and Frommer, W.B. 1992. The GUS reporter system as a tool to study plant gene expression. In: S.R. Gallagher (Ed.) GUS Protocols, Academic Press, San Diego, CA, pp. 23–43.
Mason, S.W., Sander, E.E., Evers, R. and Grummt, I. 1998. Termination of mammalian RNA polymerase I transcription. In: M.R. Paule (Ed.) Transcription of Eukaryotic Ribosomal RNA Genes by RNA Polymerase I, Springer-Verlag, Berlin, pp. 179–194.
Mélèse, T. and Xue, Z. 1995. The nucleolus: an organelle formed by the act of building a ribosome. Curr. Opin. Cell Biol. 7: 319–324.
Morrow, B.E., Johnson, S.P. and Warner, J.R. 1993. The rRNA enhancer regulates rRNA transcription in Saccharomyces cerevisiae. Mol. Cell. Biol. 13: 1283–1289.
Moss, T. and Stefanovsky, V. 1995. Promotion and regulation of ribosomal transcription in eukaryotes by RNA polymerase I. Prog. Nucl. Acids Res. Mol. Biol. 50: 25–66.
Nawrath, C., Schell, J. and Koncz, C. 1990 Homologous domains of the largest subunit of eucaryotic RNA polymerase II are conserved in plants. Mol. Gen. Genet. 223: 65–75.
Olson, M.O.J., Dundr, M. and Szebeni, A. 2000. The nucleolus: and old factory with unexpected capabilities. Trends Cell Biol. 10: 189–196.
Ott, R. and Hansen, L.K. 1996. Repeated sequences from the Arabidopsis thaliana genome function as enhancers in transgenic tobacco. Mol. Gen. Genet. 252: 563–571.
Paule, M.R. 1998. Transcription of Ribosomal RNA Genes by Eukaryotic RNA Polymerase I. Springer-Verlag, Berlin.
Pikaard, C.S. 2000 The epigenetics of nucleolar dominance. Trends Genet. 16: 495–500.
Shaw, PJ. and Jordan, E.G. 1995. The nucleolus. Annu. Rev. Cell Dev. Biol. 11: 93–121.
Sternberg, N., Sauer, B., Hoess, R. and Abremski, K. 1986. Bacteriophage P1 cre gene and its regulatory region. Evidence for multiple promoters and for regulation by DNA methylation. J. Mol. Biol. 187: 197–212.
Swoboda, P., Gal, S., Hohn, B. and Puchta, H. 1994. Intrachromosomal homologous recombination in whole plants. EMBO J. 13: 484–489.
Tinland, B., Hohn, B. and Puchta, H. 1994. Agrobacterium tumefaciens transfers single-stranded transferred DNA (T-DNA) into the plant cell nucleus. Proc. Natl. Acad. Sci. USA 91: 8000–8004.
Ñlker, B., Allen, G.C., Thompson, W.F., Spiker, S. and Weissinger, A.K. 1999. A tobacco matrix attachment region reduces the loss of transgene expression in the progeny of transgenic tobacco plants. Plant J. 18: 253–263.
Vain, P., Worland, B., Kohli, A., Snape, J.W., Christou, P., Allen, G.C. and Thompson, W.F. 1999. Matrix attachment regions increase transgene expression levels and stability in transgenic rice plants and their progeny. Plant J. 18: 233–242.
Valvekens, D., Van Montagu, M. and Van Lijsebettens, M. 1988. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc. Natl. Acad. Sci. USA 85: 5536–5540.
Wai, H., Johzuka, K., Vu, L., Eliason, K., Kobayashi, T., Horiuchi, T. and Nomura, M. 2001. Yeast RNA polymerase I enhancer is dispensable for transcription of the chromosomal rRNA gene and cell growth, and its apparent transcription enhancement from ectopic promoters requires Fob1 protein. Mol. Cell. Biol. 21: 5541–5553.
Wanzenböck, E.-M., Schöfer, C., Schweizer, D. and Bachmair, A. 1997. Ribosomal transcription units integrated via T-DNA transformation associate with the nucleolus and do not require upstream repeat sequences for activity in Arabidopsis thaliana. Plant J. 11: 1007–1016.
Wilson, K.J., Jefferson, R.A. and Hughes, S.G. 1992. The Escherichia coli gus operon: introduction and expression of the gus operon in E. coli and the occurrence and use of GUS in other bacteria. In: S.R. Gallagher (Ed.) GUS Protocols, Academic Press, San Diego, CA, pp. 7–22.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schlögelhofer, P., Nizhynska, V., Feik, N. et al. The upstream Sal repeat-containing segment of Arabidopsis thaliana ribosomal DNA intergenic region (IGR) enhances the activity of adjacent protein-coding genes. Plant Mol Biol 49, 653–663 (2002). https://doi.org/10.1023/A:1015556531074
Issue Date:
DOI: https://doi.org/10.1023/A:1015556531074