Abstract
Matrix attachment regions (MARs) are binding sites for nuclear scaffold proteins in vitro, and are proposed to mediate the attachment of chromatin to the nuclear scaffold in vivo. Previous reports suggest that MAR elements may stabilize transgene expression. Here, we tested the effects of two maize MAR elements (P-MAR from the P1-rr gene, and Adh1-MAR from the adh1 gene) on the expression of a gusA reporter gene driven by three different promoters: the maize p1 gene promoter, a wheat peroxidase (WP) gene promoter, or a synthetic promoter (Rsyn7). The inclusion of P-MAR or Adh1-MAR on P::GUS transgene constructs did not reduce variation in the levels of GUS activity among independent transformation events, nor among the progeny derived from each event. The Adh1-MAR element did not affect GUS expression driven by the WP promoter, but did modify the spatial pattern of expression of the Rsyn7::GUS transgene. These results indicate that, in transgenic maize plants, the effects of MAR elements can vary significantly depending upon the promoter used to drive the transgene.
Article PDF
Similar content being viewed by others
References
Allen GC, Hall GE, Childs LC, Weissinger AK, Spiker S and Thompson WF (1993) Scaffold attachment regions increase gene expression in stably transformed plant cells. Plant Cell 5: 603–613.
Allen GC, Hall Jr G, Michalowski S, Newman W, Spiker S, Weissinger AK et al. (1996) High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco. Plant Cell 8: 899–913.
Allen GC, Spiker S and Thompson WF (2000) Use of matrix attachment regions (MARs) to minimize transgene silencing. Plant Mol Biol 43: 361–376.
An G, Mitra A, Choi H, Costa M, Thornburg R and Ryan C (1989) Functional analysis of the 3 control region of potato wound inducible proteinase inhibitor II gene. Plant Cell 1: 115–122.
Armstrong CL and Green CE (1985) Establishment and maintenance of friable, embryogenic maize callus and the involvement of L-proline. Planta 164: 207–214.
Avramova Z and Bennetzen JL (1993) Isolation of matrices from maize leaf nuclei: identification of matrix-binding site adjacent to the adh1 gene. Plant Mol Biol 22: 1135–1143.
Avramova Z, SanMiguel P, Georgieva E and Bennetzen JL (1995) Matrix attachment regions and transcribed sequences within a long chromosomal continuum containing maize adh1. Plant Cell 7: 1667–1680.
Avramova Z, Tichonov A, Chen M and Bennetzen JL (1998) Matrix attachment regions and structural colinearity in the genomes of two grass species. Nucl Acids Res 3: 761–767.
Baulcombe DC and English JJ (1996) Ectopic pairing of homologous DNA and posttranscriptional gene silencing in transgenic plants. Curr Biotech 7: 173–180.
Berezney R and Coffey D (1974) Identification of a nuclear protein matrix. Biochem Biophys Res Comm 60: 1410–1419.
Bonifer C, Yannoutsos N, Kruger G, Grosveld F and Sippel AE (1994) Dissection of the locus control function located on the chicken lysozyme gene domain in transgenic mice. Nucl Acids Res 22: 4202–4210.
Bowen B (1992) Anthocyanin genes as visual markers in transformed maize tissues. In Gallagher SR (ed.), GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression. (pp. 163–177) Academic Press, San Diego, California.
Breyne P, van Montagu M, Depicker A and Gheysen G (1992) Characterization of a plant scaffold attachment region in a DNA fragment that normalizes transgene expression in tobacco. Plant Cell 4: 463–471.
Brouwer CR (1998) Epigenetic effects of matrix attachment regions on transgene expression in maize callus line. Dissertation, Iowa State University, pp. 133.
Bruce WB and Sims L (1997) Synthetic Plant Core promoter and Upstream Regulatory Element. PCT patent application WO 97/47756.
Bureau TE and Wessler SR (1992) Tourist: a large family of small inverted repeat elements frequently associated with maize genes. Plant Cell 4: 1283–1294.
Campbell JL and Newton CS (1991) In: Broach JR, Pringe JR and Jones EW (eds), TheMolecular and Cellular Biology of the Yeast Saccharomyces: Genome Dynamics, Protein Sythesis and Energetics. (pp. 41–146) Cold Spring Harbor Press, Cold Spring Harbor, N.Y.
Christensen AH and Quail PH (1996) Ubiquitin promoter-based vectors for high level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res 5: 213–218.
Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY et al. (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18: 659–668.
Cocciolone SM, Sidorenko LV, Chopra S, Dixon PM and Peterson T (2000) Hierarchical patterns of transgene expression indicate involvement of developmental mechanisms in the regulation of the maize P1-rr promoter. Genetics 156: 839–846.
Cockerill PN and Garrard WT (1986) Chromosomal loop anchorage sites appear to be evolutionarily conserved. Cell 44: 273–282.
Conover WJ (1980) Practical Non-parametrical Statistics. 2nd edn, Willey, New York, 493 pp.
D'Ugo E, Bruni R, Argentini C, Giuseppetti R and Rapicetta M (1998) Identification of scaffold/matrix attachment region in recurrent site of woodchuck hepatitis virus integration. DNA Cell Biol 17: 519–527.
Das OP and Messing J (1994) Variegated phenotype and developmental methylation changes of a maize allele originating from epimutation. Genetics 136: 1121–1141.
De Block M, Botterman J, Vanderwiele M, Dockx J, Thoen C, Gossele V et al. (1987) Engineering herbicide resistance in plants by expression of detoxifying enzyme. EMBO J 6: 2713–2518.
de Sepulveda P, Salaun P, Maas N, Andre C and Panthier J (1995) SARs do not impair position-dependent expression of a kit/lacZ transgene. Biochem Biophys Res Comm 211: 735–741.
Dennis ES, Gerlach WL, Pryor AJ, Bennetzen JL, Inglis A, Llewellyn D et al. (1984) Molecular analysis of the alcohol dehydrogenase (adh1) gene of maize. Nucl Acids Res 12: 3983–4000.
Devereux J, Haeberli P and Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395.
Gallie DR, Sleat DE,Watts JW, Turner PC and Wilson TMA (1987) The 5 leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucl Acids Res 15: 3257–3273.
Gasser SM and Laemmli UK (1986) Cohabitation of scaffold binding regions with upstream/enhancer elements of three developmentally regulated genes of D. melanogaster.Cell 46: 521–530.
Gasser SM, Amati BB, Cardenas ME and Hofman JF-X (1989) Studies on scaffold attachment sites and their relation to genome function. Int Rev Cytol 119: 57–96.
Gordon-Kamm WJ, Baszczynski WB, Bruce WB and Tomes DT (1999) In: Vasil IK (ed.), Transgenic Cereals-Zea mays (maize). Molecular Improvement of Cereal Crops. (pp. 189–253) Kluwer Academic Publishers.
Gray S and Levine M (1996) Transcriptional repression in development. Curr Op Cell Biol 8: 358–364.
Grotewold E, Athma P and Peterson T (1991) Alternatively spliced products of the maize P gene encode proteins with homology to the DNA binding domain of MYB-like transcriptional factor. Proc Natl Acad Sci USA 88: 4587–4591.
Hertig C, Rebmann G, Bull J, Mauch F and Dudler R (1991) Sequence and tissue specific expression of a putative peroxidase gene from wheat (Triticum aestivum L.). Plant Mol Biol 16: 171–174.
Huber MC, Bosch FX, Sippel AE and Bonifer C (1994) Chromosomal position effect in chicken lysozyme gene transgenic mice are correlated with suppression of DNase I hypersensitive site formation. Nucl Acids Res 22: 4195–4201.
Jackson DA (1991) Structure-function relationships in eukaryotic nuclei. Bioassays 13: 1–10.
Jefferson R (1986) Beta-glucoronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 83: 8447–8451.
Jefferson R (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405.
Kaiserman HB, Marini NJ, Poll EHA and Benbow R (1990) In: Strauss PPR and Wilson PSH (eds), The Eukaryotic Nucleus: Molecular Biochemistry and Macromolecular Assemblies. (pp. 783–811) Caldwell, New Jersey
Kalos M and Fournier RFK (1995) Position independent transgene expression mediated by boundary elements from the alipoprotein B chromatin domain.Mol Cell Biol 15: 198–207.
Klein T, Kornstein L, Sanford J and Fromm M (1989) Genetic transformation of maize cells by particle bombardment. Plant Physiol 91: 440–444.
Kohwi-Shigematsu T, Maass K and Bode J (1997) A thymocyte factor SATB1 suppresses transcription of stably integrated matrix-attachment region-linked reporter genes. Biochemistry 36: 12005–12010.
Kramer JA, Singh GB and Krawetz SA (1996) Computer-assisted search for sites of nuclear matrix attachment. Genomics 33: 305–308.
Kumpatla SP, Teng W, Buchholz WG and Hall TC (1997) Epigenetic transcriptional silencing and 5-azacytidine-mediated reactivation of a complex transgene in rice. Plant Physiol 115: 361–373.
Lauber AH, Barrett TJ, Subramaniam M, Schuchard M and Spelberg TC (1997) A DNA-binding element for a steroid receptor-binding factor is flanked by dual nuclear matrix DNA attachment sites in the c-myc gene promoter. J Biol Chem 36: 24657–24665.
Lechelt C, Peterson T, Laird A, Chen J, Dellaporta S, Dennis E et al. (1989) Isolation and molecular analysis of the maize P locus. Mol Gen Genet 219: 225–234.
Lee TH, Kim SJ, Han YM, Yu DY, Lee CS, Choi YJ et al. (1998) Matrix attachment region sequences enhanced the expression frequency of a whey acidic protein/human lactoferritin fusion gene in the mammary gland of transgenic mice. Mol Cell 8: 530–536.
Liu J, Bramblett D, Zhu Q, Losano M, Kobayashi R, Ross SR et al. (1997) The matrix attachment region-binding protein SATB1 participates in negative regulation of tissue-specific gene expression. Mol Cell Biol 17: 5275–5287.
Liu JW and Tabe LM (1998) The influences of two plant nuclear matrix attachment regions (MARs) on gene expression in transgenic plants. Plant Cell Physiol 39: 115–123.
Makarova O, Gorneva G, Wu F, Faturin V, Villeponteau B, Poliani L et al. (1996) Incorporation of nuclear matrix regions into the herpes simplex virus type 1 genome does not induce long-term expression of a foreign gene during latency. Gene Therap 3: 829–833.
McKnight RA, Spencer M, Wall RJ and Hennighausen L (1996) Severe position effects imposed on a 1 kb mouse whey acidic protein gene promoter are overcome by heterologous matrix attachment regions. Mol Reprod Dev 2: 179–184.
Mirkovitch J, Mirault ME and Laemmli UK (1984) Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold. Cell 39: 223–232.
Mlynarova L, Loopen A, Heldens J, Jansen RC, Keizer P, Stiekema WJ et al. (1994) Reduced position effect in mature transgenic plants conferred by the chicken lysozyme matrix-associated region. Plant Cell 6: 417–426.
Moreno MA, Chen 6J, Greenblatt I and Dellaporta SL (1992) Reconstitutional mutagenesis of the maize P-vv allele by short-range Ac transpositions. Genetics 131: 939–956.
Muller BF, Paulsen D and Deppert W (1996) Specific binding of MAR/SAR DNA-elements by mutant p53. Oncogene 12: 1941–1952.
Murashige T and Scoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.
Nabirochkin S, Ossokina M and Heidmann T (1998) A nuclear matrix/scaffold attachment region co-localizes with the gypsy retrotransposon insulator sequence. J Biol Chem 273: 2473–2479.
Oancea AE, Berru M and Shulman MJ (1997) Expression of the (recombinant) endogenous immunoglobulin heavy-chain locus requires the intronic matrix attachment regions. Mol Cell Biol 17: 2658–2668.
Phi-Van L and Stratling WH (1996) Dissection of the ability of the chicken lysozyme gene 5_ matrix attachment region to stimulate transgene expression and to dampen position effect. Biochemistry 35: 10735–10742.
Rao BS, Zannis-Hadjopoulos M, Price GB, Reitman M and Martin RG (1990) Sequence similarities among ori-enriched (ors) fragments. Gene 87: 233–242.
Rosner B and Grove D (1999) Use of the Mann–Whitney test for clustered data. Stat Med 18: 1387–1400.
Saghai-Maroof MA, Soliman KM, Jordensen RA and Allard RW (1984) Ribosomal DNA spacer-length polymorphism in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81: 8014–8018.
Sander Mand Hsieh TS (1985) Drosophila topoisomerase II doublestrand DNA cleavage: analysis of DNA sequence homology at cleavage site. Nucl Acid Res 13: 1057–1072.
Sidorenko LV, Li X, Tagliani L, Bowen B and Peterson T (1999) Characterization of the regulatory elements of the maize P1-rr gene by transient expression assays. Plant Mol Biol 39: 11–19.
Sidorenko LV, Li X, Cocciolone SM, Chopra S, Tagliani L, Bowen B et al. (2000) Complex structure of a maize Myb gene promoter: functional analysis in transgenic plants. Plant J 22: 1419–1425.
Sidorenko LV and Peterson T (2001) Transgene-induced silencing identifies sequences involved in the establishment of paramutation of the maize p1 gene. Plant Cell 13: 319–335.
Singh GB, Kramer JA and Krawetz SA (1997) Mathematical model to predict regions of chromatin attachment to the nuclear matrix. Nucl Acids Res 25: 1419–1425.
Stam M, Mol JNM and Kooter JM (1997) The silence of genes in transgenic plants. Ann Bot 79: 3–12.
Suzuki M, Izuta S, Savoysky E, Sakurai T, Simbulan C, Tatebe M et al. (1993) Deoxypyrimidine cluster mediates the priming by calf thymus DNA primase subunit. Biochem Mol Biol Int 29: 645–652.
Thompson E, Christians E, Stinnakre MG and Renard J-P (1994) Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues. Mol Cell Biol 14: 4694–4703.
Tikhonov AP, Bennetzen JL and Avramova ZV (2000) Structural domains and matrix attachment regions along colinear chromosomal segments of maize and sorghum. Plant Cel 12: 249–264.
Ulker B, Allen GC, Thompson WF, Spiker S and Weissinger AK (1999) A tobacco matrix attachment region reduces the loss of transgene expression in the progeny of transgenic tobacco. Plant J 18: 253–263.
Vain P,Worland B, Kohli A, Snape JW, Christou P, Allen GC (1999) Matrix attachment region increase transgene expression levels and stability in transgenic rice plants and their progeny. Plant J 18: 233–242.
van der Geest AH and Hall TC (1997) The beta-phaseolin 5_ matrix attachment region acts as an enhancer facilitator. Plant Mol Biol 33: 553–557.
van der Geest AH, Hall Jr CE, Spiker S and Hall TC (1994) The beta-phaseolin gene is flanked by matrix attachment regions. Plant J 6: 413–423.
Vaucheret H, Elmayan T, Thierry D, van der Geest A, Hall T, Conner AJ et al. (1998) Flanking matrix attachment regions (MARs) from chicken, bean, yeast or tobacco do not prevent homologydependent trans-silencing in transgene tobacco plants. Mol Gen Genet 259: 388–392.
Wise RP and Schnable PS (1994) Mapping complementary genes in maize: positioning rf1 and rf2 nuclear fertility restorer loci of Texas (T) cytoplasm relative to RFLP and visible markers. Theor Appl Genet 88: 785–795.
Wohlleben W, Arnold W, Broer I, Hillemann D, Strauch E and Puhler A (1988) Nucleotide sequence of the phosphinothricin N-acetyltransferase from Streptomyces viridochromogenes TN494 and its expression in Nicotiana tabaccum. Gene 70: 25–37.
Zhang P, Chopra S and Peterson T (2000) A segmental duplication generated differentially expressed myb-homologous genes in maize. Plant Cell 12: 2311–2322.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sidorenko, L., Bruce, W., Maddock, S. et al. Functional Analysis of Two Matrix Attachment Region (MAR) Elements in Transgenic Maize Plants. Transgenic Res 12, 137–154 (2003). https://doi.org/10.1023/A:1022908614356
Issue Date:
DOI: https://doi.org/10.1023/A:1022908614356