The gene encoding Arabidopsis acyl-CoA-binding protein 3 is pathogen inducible and subject to circadian regulation
In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation...
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| Published in | Journal of experimental botany Vol. 63; no. 8; pp. 2985 - 3000 |
|---|---|
| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford
Oxford University Press
01.05.2012
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| Subjects | |
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| Abstract | In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation of ACBP3, a 1.7 kb 5'-flanking region of ACBP3 and its deletion derivatives were characterized using β-glucuronidase (GUS) fusions. A 374 bp minimal fragment (-151/+223) could drive GUS expression while a 1698 bp fragment (-1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic Arabidopsis harbouring the largest (1698 bp) ACBP3pro::GUS fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized ACBP3 functions in plant defence. A 160 bp region (-434/-274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, -341/-338) interacted specifically with leaf nuclear proteins from dark-treated Arabidopsis, while GT-1 (-406/-401) binds both dark- and light-treated Arabidopsis, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of ACBP3. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (-543/-434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (-516/-512) was identified to bind nuclear proteins from pathogen-infected Arabidopsis leaves, providing the basis for pathogen-inducible regulation of ACBP3 expression. Thus, three cis-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of ACBP3 are proven functional in the regulation of ACBP3. |
|---|---|
| AbstractList | In
Arabidopsis thaliana
, acyl-CoA-binding protein 3 (
ACBP3
), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to
Pseudomonas syringae
pv.
tomato
DC3000. To understand the regulation of
ACBP3
, a 1.7 kb 5'-flanking region of
ACBP3
and its deletion derivatives were characterized using β-glucuronidase (GUS) fusions. A 374 bp minimal fragment (–151/+223) could drive GUS expression while a 1698 bp fragment (–1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic
Arabidopsis
harbouring the largest (1698 bp)
ACBP3pro::GUS
fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized
ACBP3
functions in plant defence. A 160 bp region (–434/–274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, –341/–338) interacted specifically with leaf nuclear proteins from dark-treated
Arabidopsis
, while GT-1 (–406/–401) binds both dark- and light-treated
Arabidopsis
, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of
ACBP3
. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (–543/–434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (–516/–512) was identified to bind nuclear proteins from pathogen-infected
Arabidopsis
leaves, providing the basis for pathogen-inducible regulation of
ACBP3
expression. Thus, three
cis
-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of
ACBP3
are proven functional in the regulation of
ACBP3
. In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation of ACBP3, a 1.7 kb 5'-flanking region of ACBP3 and its deletion derivatives were characterized using β-glucuronidase (GUS) fusions. A 374 bp minimal fragment (-151/+223) could drive GUS expression while a 1698 bp fragment (-1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic Arabidopsis harbouring the largest (1698 bp) ACBP3pro::GUS fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized ACBP3 functions in plant defence. A 160 bp region (-434/-274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, -341/-338) interacted specifically with leaf nuclear proteins from dark-treated Arabidopsis, while GT-1 (-406/-401) binds both dark- and light-treated Arabidopsis, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of ACBP3. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (-543/-434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (-516/-512) was identified to bind nuclear proteins from pathogen-infected Arabidopsis leaves, providing the basis for pathogen-inducible regulation of ACBP3 expression. Thus, three cis-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of ACBP3 are proven functional in the regulation of ACBP3.In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation of ACBP3, a 1.7 kb 5'-flanking region of ACBP3 and its deletion derivatives were characterized using β-glucuronidase (GUS) fusions. A 374 bp minimal fragment (-151/+223) could drive GUS expression while a 1698 bp fragment (-1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic Arabidopsis harbouring the largest (1698 bp) ACBP3pro::GUS fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized ACBP3 functions in plant defence. A 160 bp region (-434/-274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, -341/-338) interacted specifically with leaf nuclear proteins from dark-treated Arabidopsis, while GT-1 (-406/-401) binds both dark- and light-treated Arabidopsis, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of ACBP3. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (-543/-434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (-516/-512) was identified to bind nuclear proteins from pathogen-infected Arabidopsis leaves, providing the basis for pathogen-inducible regulation of ACBP3 expression. Thus, three cis-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of ACBP3 are proven functional in the regulation of ACBP3. In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation of ACBP3, a 1.7 kb 5'-flanking region of ACBP3 and its deletion derivatives were characterized using β-glucuronidase (GUS) fusions. A 374 bp minimal fragment (–151/+223) could drive GUS expression while a 1698 bp fragment (–1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic Arabidopsis harbouring the largest (1698 bp) ACBP3pro::GUS fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized ACBP3 functions in plant defence. A 160 bp region (–434/–274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, –341/–338) interacted specifically with leaf nuclear proteins from dark-treated Arabidopsis, while GT-1 (–406/–401) binds both dark- and light-treated Arabidopsis, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of ACBP3. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (–543/–434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (–516/–512) was identified to bind nuclear proteins from pathogen-infected Arabidopsis leaves, providing the basis for pathogen-inducible regulation of ACBP3 expression. Thus, three cis-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of ACBP3 are proven functional in the regulation of ACBP3. In Arabidopsis thaliana, acyl-CoA-binding protein 3 (ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It promotes autophagy-mediated leaf senescence and confers resistance to Pseudomonas syringae pv. tomato DC3000. To understand the regulation of ACBP3, a 1.7 kb 5'-flanking region of ACBP3 and its deletion derivatives were characterized using beta -glucuronidase (GUS) fusions. A 374 bp minimal fragment (-151/+223) could drive GUS expression while a 1698 bp fragment (-1475/+223) conferred maximal activity. Further, histochemical analysis on transgenic Arabidopsis harbouring the largest (1698 bp) ACBP3pro::GUS fusion displayed ubiquitous expression in floral organs and vegetative tissues (vascular bundles of leaves and stems), consistent with previous results showing that extracellularly localized ACBP3 functions in plant defence. A 160 bp region (-434/-274) induced expression in extended darkness and caused down-regulation in extended light. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay showed that the DNA-binding with one finger box (Dof-box, -341/-338) interacted specifically with leaf nuclear proteins from dark-treated Arabidopsis, while GT-1 (-406/-401) binds both dark- and light-treated Arabidopsis, suggesting that Dof and GT-1 motifs are required to mediate circadian regulation of ACBP3. Moreover, GUS staining and fluorometric measurements revealed that a 109 bp region (-543/-434) was responsive to phytohormones and pathogens. An S-box of AT-rich sequence (-516/-512) was identified to bind nuclear proteins from pathogen-infected Arabidopsis leaves, providing the basis for pathogen-inducible regulation of ACBP3 expression. Thus, three cis-responsive elements (Dof, GT-1, and the S-box) in the 5'-flanking region of ACBP3 are proven functional in the regulation of ACBP3. |
| Author | Zheng, Shu-Xiao Xiao, Shi Chye, Mee-Len |
| AuthorAffiliation | School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China |
| AuthorAffiliation_xml | – name: School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China |
| Author_xml | – sequence: 1 givenname: Shu-Xiao surname: Zheng fullname: Zheng, Shu-Xiao – sequence: 2 givenname: Shi surname: Xiao fullname: Xiao, Shi – sequence: 3 givenname: Mee-Len surname: Chye fullname: Chye, Mee-Len |
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| Issue | 8 |
| Keywords | Light effect dark/light regulation GT-1 cis-acting element Acyl Circadian rhythm Defensive response Arabidopsis thaliana Arabidopsis ACBP3 Binding protein defence response Gene Cruciferae Dicotyledones Pathogenic S-box Angiospermae Botany Regulatory sequence Spermatophyta Cis effect Experimental plant Dof-box Photoregulation |
| Language | English |
| License | CC BY 4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details) |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present address: State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China. |
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| Snippet | In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain.... In Arabidopsis thaliana, acyl-CoA-binding protein 3 (ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain. It... In Arabidopsis thaliana, acyl-CoA-binding protein 3 ( ACBP3), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding domain.... In Arabidopsis thaliana , acyl-CoA-binding protein 3 ( ACBP3 ), one of six ACBPs, is unique in terms of the C-terminal location of its acyl-CoA-binding... |
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| SubjectTerms | 5' Flanking Region 5' Flanking Region - genetics Arabidopsis Arabidopsis - drug effects Arabidopsis - genetics Arabidopsis - growth & development Arabidopsis - microbiology Arabidopsis Proteins Arabidopsis Proteins - genetics Arabidopsis Proteins - metabolism Arabidopsis thaliana Base Sequence binding proteins Biological and medical sciences Carrier Proteins Carrier Proteins - genetics Carrier Proteins - metabolism Circadian Rhythm Circadian Rhythm - drug effects Circadian Rhythm - genetics Darkness deoxyribonuclease I Deoxyribonuclease I - metabolism DNA Footprinting drug effects electrophoresis Electrophoretic Mobility Shift Assay Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Developmental Gene Expression Regulation, Developmental - drug effects Gene Expression Regulation, Plant Gene Expression Regulation, Plant - drug effects genes Genes, Plant Genes, Plant - genetics genetics Glucuronidase Glucuronidase - metabolism growth & development leaf protein leaves Lycopersicon esculentum metabolism microbiology Molecular Sequence Data pathogens pharmacology physiology Plant Growth Regulators Plant Growth Regulators - pharmacology Plant physiology and development Pseudomonas syringae Pseudomonas syringae - drug effects Pseudomonas syringae - physiology Pseudomonas syringae pv. tomato Recombinant Fusion Proteins Recombinant Fusion Proteins - metabolism Regulatory Sequences, Nucleic Acid Regulatory Sequences, Nucleic Acid - genetics Reproducibility of Results Research Papers Sequence Analysis, DNA Sequence Deletion Sequence Deletion - genetics stems Time Factors vascular bundles |
| Title | The gene encoding Arabidopsis acyl-CoA-binding protein 3 is pathogen inducible and subject to circadian regulation |
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