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Both the constitutive Cauliflower Mosaic Virus 35S and tissue-specific AGAMOUS enhancers activate transcription autonomously in Arabidopsis thaliana

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Abstract

The expression of eukaryotic genes from their cognate promoters is often regulated by the action of transcriptional enhancer elements that function in an orientation-independent manner either locally or at a distance within a genome. This interactive nature often provokes unexpected interference within transgenes in plants, as reflected by misexpression of the introduced gene and undesired phenotypes in transgenic lines. To gain a better understanding of the mechanism underlying enhancer/promoter interactions in a plant system, we analyzed the activation of a β-glucuronidase (GUS) reporter gene by enhancers contained within the AGAMOUS second intron (AGI) and the Cauliflower Mosaic Virus (CaMV) 35S promoter, respectively, in the presence and absence of a target promoter. Our results indicate that both the AGI and 35S enhancers, which differ significantly in their species of origin and in the pattern of expression that they induce, have the capacity to activate the expression of a nearby gene through the promoter-independent initiation of autonomous transcriptional events. Furthermore, we provide evidence that the 35S enhancer utilizes a mechanism resembling animal- and yeast-derived scanning or facilitated tracking models of long-distance enhancer action in its activation of a remote target promoter.

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References

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ et al (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657. doi:10.1126/science.1086391

    Article  PubMed  Google Scholar 

  • Banerji J, Rusconi S, Schaffner W (1981) Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. Cell 2:299–308

    Article  Google Scholar 

  • Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721

    Article  CAS  PubMed  Google Scholar 

  • Blackwood EM, Kadonaga JT (1998) Going the distance: a current view of enhancer action. Science 281:60–63. doi:10.1126/science.281.5373.60

    Article  CAS  PubMed  Google Scholar 

  • Bondarenko VA, Liu YV, Jiang YI, Studitsky VM (2003) Communication over a large distance: enhancers and insulators. Biochem Cell Biol 81:241–251. doi:10.1139/O03-051

    Article  CAS  PubMed  Google Scholar 

  • Broders F, Scherrer K (1987) Transcription of the alpha globin gene domain in normal and AEV-transformed chicken erythroblasts: mapping of giant globin-specific RNA including embryonic and adult genes. Mol Gen Genet 209:210–220. doi:10.1007/BF00329645

    Article  CAS  PubMed  Google Scholar 

  • Bulger M, Groudine M (1999) Looping versus linking: toward a model for long-distance gene activation. Genes Dev 13:2465–2477

    Article  CAS  PubMed  Google Scholar 

  • Busch MA, Bomblies K, Weigel D (1999) Activation of a floral homeotic gene in Arabidopsis. Science 285:585–587. doi:10.1126/science.285.5427.585

    Article  CAS  PubMed  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  CAS  PubMed  Google Scholar 

  • Deyholos MK, Sieburth LE (2000) Separable whorl-specific expression and negative regulation by enhancer elements within the AGAMOUS second intron. Plant Cell 12:1799–1810

    Article  CAS  PubMed  Google Scholar 

  • Dobi KC, Winston F (2007) Analysis of transcriptional activation at a distance in Saccharomyces cerevisiae. Mol Cell Biol 27:5575–5586. doi:10.1128/MCB.00459-07

    Article  CAS  PubMed  Google Scholar 

  • Donoghue M, Ernst H, Wentworth B, Nadal-Ginard B, Rosenthal N (1988) A muscle-specific enhancer is located at the 3′ end of the myosin light-chain 1/3 gene locus. Genes Dev 2:1779–1790. doi:10.1101/gad.2.12b.1779

    Article  CAS  PubMed  Google Scholar 

  • Dorsett D (1999) Distant liaisons: long-range enhancer–promoter interactions in Drosophila. Curr Opin Genet Dev 9:505–514. doi:10.1016/S0959-437X(99)00002-7

    Article  CAS  PubMed  Google Scholar 

  • Drewell RA, Bae E, Burr J, Lewis EB (2002) Transcription defines the embryonic domains of cis-regulatory activity at the Drosophila bithorax complex. Proc Natl Acad Sci USA 99:16853–16858. doi:10.1073/pnas.222671199

    Article  CAS  PubMed  Google Scholar 

  • Emerson BM, Nickol JM, Jackson PD, Felsenfeld G (1987) Analysis of the tissue-specific enhancer at the 3′ end of the chicken adult β-globin gene. Proc Natl Acad Sci USA 84:4786–4790

    Article  CAS  PubMed  Google Scholar 

  • Fiering S, Whitelaw E, Martin DIK (2000) To be or not to be active: the stochastic nature of enhancer action. Bioessays 22:381–387

    Article  CAS  PubMed  Google Scholar 

  • Gillies SD, Morrison SL, Oi VT, Tonegawa S (1983) A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobin heavy chain gene. Cell 33:717–728

    Article  CAS  PubMed  Google Scholar 

  • Hatzis P, Talianidis I (2002) Dynamics of enhancer–promoter communication during differentiation-induced gene activation. Mol Cell 10:1467–1477. doi:10.1016/1097-2765(02)00786-4

    Article  CAS  PubMed  Google Scholar 

  • Hily JM, Liu Z (2009) A simple and sensitive high-throughput GFP screening in woody and herbaceous plants. Plant Cell Rep 28:493–501. doi:10.1007/s00299-008-0657-z

    Article  CAS  PubMed  Google Scholar 

  • Hily JM, Singer SD, Yang Y, Liu Z (2009) A transformation booster sequence (TBS) from Petunia hybrida functions as an enhancer-blocking insulator in Arabidopsis thaliana. Plant Cell Rep 28:1095–1104. doi:10.1007/s00299-009-0700-8

    Article  CAS  PubMed  Google Scholar 

  • Hinz U, Wolk A, Renkawitz-Pohl R (1992) Ultrabithorax is a regulator of β3 tubulin expression in the Drosophila visceral mesoderm. Development 116:543–554

    CAS  PubMed  Google Scholar 

  • Hirner B, Fischer WN, Rentsch D, Kwart M, Frommer WB (1998) Developmental control of H+/amino acid permease gene expression during seed development of Arabidopsis. Plant J 14:535–544. doi:10.1046/j.1365-313X.1998.00151.x

    Article  CAS  PubMed  Google Scholar 

  • Ho Y, Elefant F, Liebhaber SA, Cooke NE (2006) Locus control region transcription plays an active role in long-range gene activation. Mol Cell 23:365–375. doi:10.1016/j.molcel.2006.05.041

    Article  CAS  PubMed  Google Scholar 

  • Jack J, Dorsett D, DeLotto Y, Liu S (1991) Expression of the cut locus in the Drosophila wing margin is required for cell type specification and is regulated by a distant enhancer. Development 113:735–747

    CAS  PubMed  Google Scholar 

  • Jacquemin P, Oury C, Peers B, Morin A, Belayew A, Martial JA (1994) Characterization of a single strong tissue-specific enhancer downstream from the three human genes encoding placental lactogen. Mol Cell Biol 14:93–103

    CAS  PubMed  Google Scholar 

  • Jagannath A, Bandyopadhyay P, Arumugam N, Gupta V, Kumar P, Pental D (2001) The use of a Spacer DNA fragment insulates the tissue-specific expression of a cytotoxic gene (barnase) and allow high-frequency generation of transgenic male sterile lines in Brassica juncea L. Mol Breeding 8:11–23

    Article  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    CAS  PubMed  Google Scholar 

  • Kadauke S, Blobel GA (2009) Chromatin loops in gene regulation. Biochim Biophys Acta 1789:17–25. doi:10.1016:/j.bbagrm.2008.07.002

    CAS  PubMed  Google Scholar 

  • Kay R, Chan A, Daly M, McPherson J (1987) Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302. doi:10.1126/science.236.4806.1299

    Article  CAS  PubMed  Google Scholar 

  • Keaveney M, Struhl K (1998) Activator-mediated recruitment of the RNA polymerase II machinery is the predominant mechanism for transcriptional activation in yeast. Mol Cell 1:917–924. doi:10.1016/S1097-2765(00)80091-X

    Article  CAS  PubMed  Google Scholar 

  • Kim A, Zhao H, Ifrim I, Dean A (2007) β-globin intergenic transcription and histone acetylation dependent on an enhancer. Mol Cell Biol 27:2980–2986. doi:10.1128/MCB.02337-06

    Article  CAS  PubMed  Google Scholar 

  • Lescot M, Dehais P, Moreau Y, De Moor B, Rouze P, Rombauts S (2002) PlantCARE: a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327

    Article  CAS  PubMed  Google Scholar 

  • Ling J, Ainol L, Zhang L, Yu X, Pi W, Tuan D (2004) HS2 enhancer function is blocked by a transcriptional terminator inserted between the enhancer and the promoter. J Biol Chem 279:51704–51713. doi:10.1074/jbc.M404039200

    Article  CAS  PubMed  Google Scholar 

  • Ling J, Baibakov B, Pi W, Emerson BM, Tuan D (2005) The HS2 enhancer of the β-globin locus control region initiates synthesis of non-coding, polyadenylated RNAs independent of a cis-linked globin promoter. J Mol Biol 350:883–896. doi:10.1016/j.jmb.2005.05.039

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, Liu Z (2008) The second intron of AGAMOUS drives carpel- and stamen-specific expression sufficient to induce complete sterility in Arabidopsis. Plant Cell Rep 27:855–863. doi:10.1007/s00299-008-0511-3

    Article  PubMed  Google Scholar 

  • Louie MC, Yang HQ, Ma AH, Xu W, Zou JX, Kung HJ, Chen HW (2003) Androgen-induced recruitment of RNA polymerase II to a nuclear receptor-p160 coactivator complex. Proc Natl Acad Sci USA 100:2226–2230. doi:10.1073/pnas.0437824100

    Article  CAS  PubMed  Google Scholar 

  • Masternak K, Peyraud N, Krawczyk M, Barras E, Reith W (2003) Chromatin remodeling and extragenic transcription at the MHC class II locus control region. Nat Immunol 4:132–137. doi:10.1038/ni883

    Article  CAS  PubMed  Google Scholar 

  • Morris JR, Chen J-I, Geyer PK, Wu C-T (1998) Two modes of transvection: enhancer action in trans and bypass of a chromatin insulator in cis. Proc Natl Acad Sci USA 95:10740–10745

    Article  CAS  PubMed  Google Scholar 

  • Müller H-P, Matthias P, Schaffner W (1990) A transcriptional terminator between enhancer and promoter does not affect remote transcriptional control. Somat Cell Mol Genet 16:351–360. doi:10.1007/BF01232463

    Article  PubMed  Google Scholar 

  • Ptashne M, Gann A (1997) Transcriptional activation by recruitment. Nature 386:569–577. doi:10.1038/386569a0

    Article  CAS  PubMed  Google Scholar 

  • Razin S, Rynditch A, Borunova V, Ioudinkova E, Smalko V, Scherrer K (2004) The 33 kb transcript of the chicken α-globin gene domain is part of the nuclear matrix. J Cell Biochem 92:445–457. doi:10.1002/jcb.20066

    Article  CAS  PubMed  Google Scholar 

  • Rippe K, von Hippel PH, Langowski J (1995) Action at a distance: DNA-looping and initiation of transcription. Trends Biochem Sci 20:500–506. doi:10.1016/S0968-0004(00)89117-3

    Article  CAS  PubMed  Google Scholar 

  • Rogan DF, Cousins DJ, Santangelo S, Ioannou PA, Antoniou M, Lee TH, Staynov DZ (2004) Analysis of intergenic transcription in the human IL-4/IL-13 gene cluster. Proc Natl Acad Sci USA 101:2446–2451. doi:10.1073/pnas.0308327100

    Article  CAS  PubMed  Google Scholar 

  • Rosenthal N, Berglund EB, Wentworth BM, Donoghue M, Winter B, Bober E, Braun T, Arnold H-H (1990) A highly conserved enhancer downstream of the human MLC1/3 locus is a target for multiple myogenic determination factors. Nucleic Acids Res 18:6239–6246

    Article  CAS  PubMed  Google Scholar 

  • Rosso MG, Li Y, Strizhov N, Reiss B, Dekker K, Weisshaar B (2003) An Arabidopsis thaliana T-DNA mutagenized population (GABI-Kat) for flanking sequence tag-based reverse genetics. Plant Mol Biol 53:247–259. doi:10.1023/B:PLAN.0000009297.37235.4a

    Article  CAS  PubMed  Google Scholar 

  • Routledge SJE, Proudfoot NJ (2002) Definition of transcriptional promoters in the human β-globin locus control region. J Mol Biol 323:601–611. doi:10.1016/S0022-2836(02)01011-2

    Article  CAS  PubMed  Google Scholar 

  • Savidge B, Rounsley SD, Yanofsky MF (1995) Temporal relationship between the transcription of two Arabidopsis MADS box genes and the floral organ identity genes. Plant Cell 7:721–733

    Article  CAS  PubMed  Google Scholar 

  • Shang Y, Myers M, Brown M (2002) Formation of the androgen receptor transcription complex. Mol Cell 9:601–610. doi:10.1016/S1097-2765(03)00237-5

    Article  CAS  PubMed  Google Scholar 

  • Sieburth LE, Meyerowitz EM (1997) Molecular dissection of the AGAMOUS control region shows that cis elements for spatial regulation are located intragenically. Plant Cell 9:355–365

    Article  CAS  PubMed  Google Scholar 

  • Singer SD, Hily J-M, Liu Z (2010) A 1 kb bacteriophage lambda fragment functions as an insulator to effectively block enhancer–promoter interactions in Arabidopsis thaliana. Plant Mol Biol Rep 28:69–76. doi:10.1007/s11105-009-0122-3

    Article  CAS  Google Scholar 

  • Smith DL, Fedoroff NV (1995) LRP1, a gene expressed in lateral and adventitious root primordia of Arabidopsis. Plant Cell 7:735–745

    Article  CAS  PubMed  Google Scholar 

  • Spicuglia S, Kumar S, Yeh JH, Vachez E, Chasson L, Gorbatch S, Cautres J, Ferrier P (2002) Promoter activation by enhancer-dependent and-independent loading of activator and coactivator complexes. Mol Cell 10:1479–1487. doi:10.1016/S1097-2765(02)00791-8

    Article  CAS  PubMed  Google Scholar 

  • Sutherland HGE, Martin DIK, Whitelaw E (1997) A globin enhancer acts by increasing the proportion of erythrocytes expressing a linked transgene. Mol Cell Biol 17:1607–1614

    CAS  PubMed  Google Scholar 

  • Tchurikov NA, Kretova OV, Moiseeva ED, Sosin DV (2009) Evidence for RNA synthesis in the intergenic region between enhancer and promoter and its inhibition by insulators in Drosophila melanogaster. Nucleic Acids Res 37:111–122. doi:10.1093/nar/gkn926

    Article  CAS  PubMed  Google Scholar 

  • Tolhuis B, Palstra RJ, Splinter E, Grosveld F, de Laat W (2002) Looping and interaction between hypersensitive sites in the active beta-globin locus. Mol Cell 10:1453–1465. doi:10.1016/S1097-2765(02)00781-5

    Article  CAS  PubMed  Google Scholar 

  • Travers A (1999) Chromatin modification by DNA tracking. Proc Natl Acad Sci USA 96:13634–13637

    Article  CAS  PubMed  Google Scholar 

  • Tuan D, Kong S, Hu K (1992) Transcription of the hypersensitive site HS2 enhancer in erythroid cells. Proc Natl Acad Sci USA 89:11219–11223

    Article  CAS  PubMed  Google Scholar 

  • Walters MC, Fiering S, Eidemiller J, Magis W, Groudine M, Martin DIK (1995) Enhancers increase the probability but not the level of gene expression. Proc Natl Acad Sci USA 92:7125–7129

    Article  CAS  PubMed  Google Scholar 

  • Wang Q, Carroll J, Brown M (2005) Spatial and temporal recruitment of androgen receptor and its coactivators involves chromosomal looping and polymerase tracking. Mol Cell 19:631–642. doi:10:1016/j.molcel.2005.07.018

    Article  CAS  PubMed  Google Scholar 

  • Zheng X, Deng W, Luo K, Duan H, Chen Y, McAvoy R, Song S, Pei Y, Li Y (2007) The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters. Plant Cell Rep 26:1195–1203. doi:10.1007/s00299-007-0307-x

    Article  CAS  PubMed  Google Scholar 

  • Zhu X, Ling J, Zhang L, Pi W, Wu M, Tuan D (2007) A facilitated tracking and transcription mechanism of long-range enhancer function. Nucleic Acids Res 35:5532–5544. doi:10.1093/nar/gkm595

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors wish to thank Mr. Dennis Bennett (USDA-ARS, Kearneysville, WV) for his technical assistance. This study was funded by the USDA-ARS Headquarter 2007 class of postdoctoral grants and a USDA CSREES BRAG grant (2006-03701).

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  1. Stacy D. Singer & Kerik D. Cox

    Present address: Department of Plant Pathology, New York State Agricultural Experiment Station, Cornell University, 630 West North Street, Geneva, NY, 14456, USA

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  1. USDA-ARS Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Stacy D. Singer & Zongrang Liu

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  1. Stacy D. Singer
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Correspondence to Zongrang Liu.

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Singer, S.D., Cox, K.D. & Liu, Z. Both the constitutive Cauliflower Mosaic Virus 35S and tissue-specific AGAMOUS enhancers activate transcription autonomously in Arabidopsis thaliana . Plant Mol Biol 74, 293–305 (2010). https://doi.org/10.1007/s11103-010-9673-9

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