title: A cis-regulatory sequence from a short intergenic region gives rise to a strong microbe-associated molecular pattern-responsive synthetic promoter.
abstract: The high gene density in Arabidopsis thaliana leaves only relatively short intergenic regions for potential cis-regulatory sequences. To learn more about the regulation of genes harbouring only very short upstream intergenic regions, this study investigates a recently identified novel microbe-associated molecular pattern (MAMP)-responsive cis-sequence located within the 101 bp long intergenic region upstream of the At1g13990 gene. It is shown that the cis-regulatory sequence is sufficient for MAMP-responsive reporter gene activity in the context of its native promoter. The 3' UTR of the upstream gene has a quantitative effect on gene expression. In context of a synthetic promoter, the cis-sequence is shown to achieve a strong increase in reporter gene activity as a monomer, dimer and tetramer. Mutation analysis of the cis-sequence determined the specific nucleotides required for gene expression activation. In transgenic A. thaliana the synthetic promoter harbouring a tetramer of the cis-sequence not only drives strong pathogen-responsive reporter gene expression but also shows a high background activity. The results of this study contribute to our understanding how genes with very short upstream intergenic regions are regulated and how these regions can serve as a source for MAMP-responsive cis-sequences for synthetic promoter design.
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Mona
authors.initials: M
authors.lastname: Lehmeyer
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Erik K R
authors.initials: EK
authors.lastname: Hanko
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Roling
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Lilian
authors.initials: L
authors.lastname: Gonzalez
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Maren
authors.initials: M
authors.lastname: Wehrs
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany. r.hehl@tu-bs.de.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Hehl
mesh_terms: D020413:3' Untranslated Regions; D017360:Arabidopsis / Q000235:genetics / Q000382:microbiology*; D021901:DNA, Intergenic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D005813:Genes, Synthetic; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000382:microbiology; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid
chemical_list: D020413:3' Untranslated Regions; D021901:DNA, Intergenic; D010940:Plant Proteins
keywords: Bioinformatics; Databases; Plant-pathogen interaction; Synthetic biology; Transgenic plants; Transient expression
references.citation: Plant Cell. 2008 Mar;20(3):768-85
references.pmid: 18334669
references.citation: Nucleic Acids Res. 2015 Jan;43(Database issue):D1003-9
references.pmid: 25414324
references.citation: BMC Genomics. 2007 Mar 08;8:67
references.pmid: 17346352
references.citation: Eukaryot Cell. 2005 Aug;4(8):1343-52
references.pmid: 16087739
references.citation: Plant Cell Environ. 2012 Oct;35(10):1787-98
references.pmid: 22489681
references.citation: Plant J. 2003 Jan;33(2):259-70
references.pmid: 12535340
references.citation: Plant J. 2003 Apr;34(2):205-16
references.pmid: 12694595
references.citation: Plant J. 1998 Dec;16(6):735-43
references.pmid: 10069079
references.citation: Nucleic Acids Res. 2012 Jan;40(Database issue):D1202-10
references.pmid: 22140109
references.citation: Plant Mol Biol. 1992 Dec;20(6):1195-7
references.pmid: 1463855
references.citation: Nature. 2000 Dec 14;408(6814):816-20
references.pmid: 11130712
references.citation: EMBO J. 1990 Sep;9(9):2945-50
references.pmid: 2390976
references.citation: Annu Rev Phytopathol. 2005;43:205-27
references.pmid: 16078883
references.citation: Nucleic Acids Res. 1985 Jul 11;13(13):4777-88
references.pmid: 4022773
references.citation: Plant Mol Biol. 2014 Jan;84(1-2):111-23
references.pmid: 23955710
references.citation: Genomics Inform. 2014 Sep;12(3):87-97
references.pmid: 25317107
references.citation: Plant Cell. 2002 Apr;14(4):749-62
references.pmid: 11971132
references.citation: Database (Oxford). 2014 Apr 10;2014(0):bau030
references.pmid: 24727366
references.citation: EMBO J. 1987 Dec 20;6(13):3901-7
references.pmid: 3327686
references.citation: Trends Biotechnol. 2005 Jun;23(6):283-90
references.pmid: 15922080
references.citation: Nucleic Acids Res. 2014 Jan;42(1):485-98
references.pmid: 24084084
references.citation: Plant Physiol. 2012 Sep;160(1):178-91
references.pmid: 22744985
references.citation: Plant J. 2000 Apr;22(1):1-8
references.pmid: 10792815
references.citation: Proc Natl Acad Sci U S A. 2014 Feb 11;111(6):2367-72
references.pmid: 24477691
references.citation: Plant Cell. 2004 Sep;16(9):2481-98
references.pmid: 15319476
references.citation: Transgenic Res. 2014 Apr;23(2):351-63
references.pmid: 24072400
references.citation: Plant Physiol. 2014 Jan;164(1):181-200
references.pmid: 24204023
references.citation: BMC Genomics. 2005 Feb 25;6:25
references.pmid: 15733318
references.citation: Nature. 2000 Dec 14;408(6814):796-815
references.pmid: 11130711
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Nucleic Acids Res. 2014 Jul;42(12):7681-93
references.pmid: 24914054
references.citation: Plant Biotechnol J. 2016 Jan;14 (1):61-71
references.pmid: 25819608
references.citation: Mol Plant Pathol. 2000 Jul 1;1(4):243-51
references.pmid: 20572971