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Ethylene Receptor Ethylene Receptor1 Domain Requirements For Ethylene Responses In Arabidopsis Seedlings, Heejung Kim, Elizabeth Helmbrecht, M. Stalins, Christina Schmitt, Nesha Patel, Chi-Kuang Wen, Wuyi Wang, Brad Binder Apr 2011

Ethylene Receptor Ethylene Receptor1 Domain Requirements For Ethylene Responses In Arabidopsis Seedlings, Heejung Kim, Elizabeth Helmbrecht, M. Stalins, Christina Schmitt, Nesha Patel, Chi-Kuang Wen, Wuyi Wang, Brad Binder

Brad M. Binder

Ethylene influences many processes in Arabidopsis (Arabidopsis thaliana) through the action of five receptor isoforms. We used high-resolution, time-lapse imaging of dark-grown Arabidopsis seedlings to better understand the roles of each isoform in the regulation of growth in air, ethylene-stimulated nutations, and growth recovery after ethylene removal. We found that ETHYLENE RECEPTOR1 (ETR1) is both necessary and sufficient for nutations. Transgene constructs in which the ETR1 promoter was used to drive expression of cDNAs for each of the five receptor isoforms were transferred into etr1-6;etr2-3;ein4-4 triple loss-of-function mutants that have constitutive growth inhibition in air, fail to nutate ...


Proteomic Responses In Arabidopsis Thaliana Seedlings Treated With Ethylene, Ruiqiang Chen, Brad Binder, Wesley Garrett, Mark Tucker, Caren Chang, Bret Cooper Dec 2010

Proteomic Responses In Arabidopsis Thaliana Seedlings Treated With Ethylene, Ruiqiang Chen, Brad Binder, Wesley Garrett, Mark Tucker, Caren Chang, Bret Cooper

Brad M. Binder

Ethylene (ET) is a volatile hormone that modulates fruit ripening, plant growth, development and stress responses. Key components of the ET-signaling pathway identified by genetic dissection in Arabidopsis thaliana include five ET receptors, the negative regulator CTR1 and the positive regulator EIN2, all of which localize to the endoplasmic reticulum. Mechanisms of signaling among these proteins are still unresolved and targets of ET responses are not fully known. So, we used mass spectrometry to identify proteins in microsomal membrane preparations from etiolated A. thaliana seedlings maintained in ambient air or treated with ET for 3 h. We compared 3814 proteins ...


Ethylene Receptors Function As Components Of High-Molecular-Mass Protein Complexes In Arabidopsis, Brad Binder, Yi-Feng Chen, Zhiyong Gao, Robert Kerris, Wuyi Wang, G. Schaller Dec 2009

Ethylene Receptors Function As Components Of High-Molecular-Mass Protein Complexes In Arabidopsis, Brad Binder, Yi-Feng Chen, Zhiyong Gao, Robert Kerris, Wuyi Wang, G. Schaller

Brad M. Binder

Background The gaseous plant hormone ethylene is perceived in Arabidopsis thaliana by a five-member receptor family composed of ETR1, ERS1, ETR2, ERS2, and EIN4. Methodology/Principal Findings Gel-filtration analysis of ethylene receptors solubilized from Arabidopsis membranes demonstrates that the receptors exist as components of high-molecular-mass protein complexes. The ERS1 protein complex exhibits an ethylene-induced change in size consistent with ligand-mediated nucleation of protein-protein interactions. Deletion analysis supports the participation of multiple domains from ETR1 in formation of the protein complex, and also demonstrates that targeting to and retention of ETR1 at the endoplasmic reticulum only requires the first 147 amino ...


Ethylene Receptors Function As Components Of High-Molecular-Mass Protein Complexes In Arabidopsis, Yi-Feng Chen, Zhiyong Gao, Robert J. Kerriss Iii, Wuyi Wang, Brad M. Binder, G. Eric Schaller Dec 2009

Ethylene Receptors Function As Components Of High-Molecular-Mass Protein Complexes In Arabidopsis, Yi-Feng Chen, Zhiyong Gao, Robert J. Kerriss Iii, Wuyi Wang, Brad M. Binder, G. Eric Schaller

Brad M. Binder

The gaseous plant hormone ethylene is perceived in Arabidopsis thaliana by a five-member receptor family composed of ETR1, ERS1, ETR2, ERS2, and EIN4. Methodology/Principal Findings Gel-filtration analysis of ethylene receptors solubilized from Arabidopsis membranes demonstrates that the receptors exist as components of high-molecular-mass protein complexes. The ERS1 protein complex exhibits an ethylene-induced change in size consistent with ligand-mediated nucleation of protein-protein interactions. Deletion analysis supports the participation of multiple domains from ETR1 in formation of the protein complex, and also demonstrates that targeting to and retention of ETR1 at the endoplasmic reticulum only requires the first 147 amino acids ...


Ethylene-Dependent And -Independent Regulation Of Abscission, Brad Binder, Sara Patterson Jan 2009

Ethylene-Dependent And -Independent Regulation Of Abscission, Brad Binder, Sara Patterson

Brad M. Binder

Purpose of review: This review focuses on recent research on the control of abscission, highlighting ethylene dependent and independent mechanisms. Findings: Abscission is the loss of organs such as fruit, leaves and floral organs. Ethylene has long been recognised as one of the most important factors controlling abscission. However, abscission is also controlled by a complex network of factors not related to ethylene, including auxin, jasmonic acid and peptides. Recent increased understandings of the nature of both ethylene-dependent and independent abscission control are expected to suggest more refined methods for abscission control in horticultural plants. Directions for future research: A ...


The Btb Ubiqutin Ligases Eto1, Eol1 And Eol2 Act Collectively To Regulate Ethylene Biosynthesis In Arabidopsis By Controlling Type-2 Acc Synthase Levels, Brad Binder, Matthew Christians, Derek Gingerich, Moureen Hansen, Joseph Kieber, Richard Vierstra Dec 2008

The Btb Ubiqutin Ligases Eto1, Eol1 And Eol2 Act Collectively To Regulate Ethylene Biosynthesis In Arabidopsis By Controlling Type-2 Acc Synthase Levels, Brad Binder, Matthew Christians, Derek Gingerich, Moureen Hansen, Joseph Kieber, Richard Vierstra

Brad M. Binder

Ethylene biosynthesis is directed by a family of 1-aminocyclopropane-1-carboxylic acid (ACC) synthases (ACS) that convert S-adenosyl-l-methionine to the immediate precursor ACC. Members of the type-2 ACS subfamily are strongly regulated by proteolysis with various signals stabilizing the proteins to increase ethylene production. In Arabidopsis, this turnover is mediated by the ubiquitin/26 S proteasome system, using a broad complex/tramtrack/bric-a-brac (BTB) E3 assembled with the ETHYLENE OVERPRODUCER 1 (ETO1) BTB protein for target recognition. Here, we show that two Arabidopsis BTB proteins closely related to ETO1, designated ETO1-like (EOL1) and EOL2, also negatively regulate ethylene synthesis via their ability ...


Heteromeric Interactions Among Ethylene Receptors Mediate Signaling In Arabidopsis, Brad Binder, Zhiyong Gao, Chi-Kuang Wen, Yi-Feng Chen, Jianhong Chang, Yi-Hsuan Chiang, Robert Kerris, Caren Chang, G. Schaller Jul 2008

Heteromeric Interactions Among Ethylene Receptors Mediate Signaling In Arabidopsis, Brad Binder, Zhiyong Gao, Chi-Kuang Wen, Yi-Feng Chen, Jianhong Chang, Yi-Hsuan Chiang, Robert Kerris, Caren Chang, G. Schaller

Brad M. Binder

The gaseous hormone ethylene is perceived in Arabidopsis by a five member receptor family that consists of the subfamily 1 receptors ETR1 and ERS1 and the subfamily 2 receptors ETR2, ERS2, and EIN4. Previous work has demonstrated that the basic functional unit for the ethylene receptor, ETR1, is a disulfide-linked homodimer. We demonstrate here that ethylene receptors isolated from Arabidopsis also interact with each other through noncovalent interactions. Evidence that ETR1 associates with other ethylene receptors was obtained by co-purification of ETR1 with tagged versions of ERS1, ETR2, ERS2, and EIN4 from Arabidopsis membrane extracts. ETR1 preferentially associated with the ...


The Ethylene Receptors: Complex Perception For A Simple Gas, Brad Binder Jun 2008

The Ethylene Receptors: Complex Perception For A Simple Gas, Brad Binder

Brad M. Binder

Ethylene is a hormone that regulates a number of physiological and developmental events in plants. This simple olefin is perceived by a family of receptors that have homology to bacterial two-component receptors. Although the ethylene receptors have been the focus of much study over the past several decades, many questions remain concerning their structure, function, and regulation. This review provides an overview on what is known about the ethylene receptors, summarizes recent research on the receptors, and presents models for receptor function and output.


Ethylene Receptor Antagonists: Strained Alkenes Are Necessary But Not Sufficient, Brad Binder, Michael Pirrung, Anthony Bleecker, Yoshihisa Inoue, Fernando Rodríguez, Norimitsu Sugawara, Takehiko Wada, Yunfan Zou Mar 2008

Ethylene Receptor Antagonists: Strained Alkenes Are Necessary But Not Sufficient, Brad Binder, Michael Pirrung, Anthony Bleecker, Yoshihisa Inoue, Fernando Rodríguez, Norimitsu Sugawara, Takehiko Wada, Yunfan Zou

Brad M. Binder

Plants use ethylene as a hormone to control many physiological processes. Ethylene perception involves its binding to an unusual copper-containing, membrane-bound receptor. Inhibitors of ethylene action are valuable to study signaling and may have practical use in horticulture. Past investigation of alkene ligands for this receptor has identified strain as the key factor in antagonism of ethylene binding and action, consistent with known trends in metal-alkene complex stability. However, in this work, this principle could not be extended to other alkenes, prompting development of the proposal that a ring-opening reaction accounts for the unusual potency of cyclopropene ethylene antagonists. Another ...


The Effects Of Group 11 Transition Metals, Including Gold, On Ethylene Binding To The Etr1 Receptor And Growth Of Arabidopsis Thaliana, Brad Binder, Fernando Rodriguez, Anthony Bleecker, Sara Patterson Sep 2007

The Effects Of Group 11 Transition Metals, Including Gold, On Ethylene Binding To The Etr1 Receptor And Growth Of Arabidopsis Thaliana, Brad Binder, Fernando Rodriguez, Anthony Bleecker, Sara Patterson

Brad M. Binder

It has been previously shown that Cu(I) and the ethylene response antagonist, Ag(I), support ethylene binding to exogenously expressed ETR1 ethylene receptors. Both are Group 11 transition metals that also include gold. We compared the effects of gold ions with those of Cu(I) and Ag(I) on ethylene binding in exogenously expressed ETR1 receptors and on ethylene growth responses in etiolated Arabidopsis seedlings. We find that gold ions also support ethylene binding but, unlike Ag(I), do not block ethylene action on plants. Instead, like Cu(I), gold ions affect seedlings independently of ethylene signaling.


The Arabidopsis Ein3 Binding F-Box Proteins Ebf1 And Ebf2 Have Distinct But Overlapping Roles In Ethylene Signaling, Brad Binder, Joseph Walker, Jennifer Gagne, Thomas Emborg, Georg Hemmann, Anthony Bleecker, Richard Vierstra Jan 2007

The Arabidopsis Ein3 Binding F-Box Proteins Ebf1 And Ebf2 Have Distinct But Overlapping Roles In Ethylene Signaling, Brad Binder, Joseph Walker, Jennifer Gagne, Thomas Emborg, Georg Hemmann, Anthony Bleecker, Richard Vierstra

Brad M. Binder

Ethylene signaling in Arabidopsis thaliana converges on the ETHYLENE-INSENSITIVE3 (EIN3)/EIN3-Like (EIL) transcription factors to induce various responses. EIN3 BINDING F-BOX1 (EBF1) and EBF2 were recently shown to function in ethylene perception by regulating EIN3/EIL turnover. In the absence of ethylene, EIN3 and possibly other EIL proteins are targeted for ubiquitination and subsequent degradation by Cullin 1–based E3 complexes containing EBF1 and 2. Ethylene appears to block this ubiquitination, allowing EIN3/EIL levels to rise and mediate ethylene signaling. Through analysis of mutant combinations affecting accumulation of EBF1, EBF2, EIN3, and EIL1, we show that EIN3 and EIL1 ...


Rapid Kinetic Analysis Of Ethylene Growth Responses In Seedlings: New Insights Into Ethylene Signal Transduction, Brad Binder Dec 2006

Rapid Kinetic Analysis Of Ethylene Growth Responses In Seedlings: New Insights Into Ethylene Signal Transduction, Brad Binder

Brad M. Binder

Ethylene is a phytohormone that influences diverse processes in plants. Ethylene causes various changes in etiolated seedlings that differ between species and include reduced growth of shoots and roots, increased diameter of shoots, agravitropic growth, initiation of root hairs, and increased curvature of the apical hook. The inhibition of growth in etiolated seedlings has become widely used to screen for and identify mutants. This approach has led to an increased understanding of ethylene signaling. Most studies use end-point analysis after several days of exposure to assess the effects of ethylene. Recently, the use of time-lapse imaging has re-emerged as an ...


The Exoribonuclease Xrn4 Is A Component Of The Ethylene Response Pathway In Arabidopsis, Brad Binder, Thomas Potuschak, Amérin Vansiri, Esther Lechner, Richard Vierstra, Pascal Genschik Oct 2006

The Exoribonuclease Xrn4 Is A Component Of The Ethylene Response Pathway In Arabidopsis, Brad Binder, Thomas Potuschak, Amérin Vansiri, Esther Lechner, Richard Vierstra, Pascal Genschik

Brad M. Binder

EXORIBONUCLEASE4 (XRN4), the Arabidopsis thaliana homolog of yeast XRN1, is involved in the degradation of several unstable mRNAs. Although a role for XRN4 in RNA silencing of certain transgenes has been reported, xrn4 mutant plants were found to lack any apparent visible phenotype. Here, we show that XRN4 is allelic to the unidentified components of the ethylene response pathway ETHYLENE-INSENSITIVE5/ACC-INSENSITIVE1 (EIN5/AIN1) and EIN7. xrn4 mutant seedlings are ethylene-insensitive as a consequence of the upregulation of EIN3 BINDING F-BOX PROTEIN1 (EBF1) and EBF2 mRNA levels, which encode related F-box proteins involved in the turnover of EIN3 protein, a crucial ...


Ethylene-Stimulated Nutations Do Not Require Etr1 Receptor Histidine Kinase Activity, Brad Binder Oct 2006

Ethylene-Stimulated Nutations Do Not Require Etr1 Receptor Histidine Kinase Activity, Brad Binder

Brad M. Binder

Ethylene influences the growth and development of plants through the action of receptors that have homology to bacterial two-component receptors. In bacteria these receptors function via autophosphorylation of a His residue in the kinase domain followed by phosphotransfer to a conserved Asp residue in a response regulator protein. In Arabidopsis, two of the five receptor isoforms are capable of His kinase activity. However, the role of His kinase activity and phosphotransfer is unclear in ethylene signaling. A previous study showed that ethylene stimulates nutations of the hypocotyl in etiolated Arabidopsis seedlings that are dependent on the ETR1 receptor isoform. The ...


Identification Of Important Regions For Ethylene Binding And Signaling In The Transmembrane Domain Of The Etr1 Ethylene Receptor Of Arabidopsis, Brad Binder, Wuyi Wang, Jeff Esch, Shin-Han Shiu, Hasi Agula, Caren Chang, Sara Patterson, Anthony Bleecker Dec 2005

Identification Of Important Regions For Ethylene Binding And Signaling In The Transmembrane Domain Of The Etr1 Ethylene Receptor Of Arabidopsis, Brad Binder, Wuyi Wang, Jeff Esch, Shin-Han Shiu, Hasi Agula, Caren Chang, Sara Patterson, Anthony Bleecker

Brad M. Binder

The ethylene binding domain (EBD) of the Arabidopsis thaliana ETR1 receptor is modeled as three membrane-spanning helices. We surveyed ethylene binding activity in different kingdoms and performed a bioinformatic analysis of the EBD. Ethylene binding is confined to land plants, Chara, and a group of cyanobacteria but is largely absent in other organisms, consistent with our finding that EBD-like sequences are overrepresented among plant and cyanobacterial species. We made amino acid substitutions in 37 partially or completely conserved residues of the EBD and assayed their effects on ethylene binding and signaling. Mutations primarily in residues in Helices I and II ...


Ethylene Stimulates Nutations That Are Dependent On The Etr1 Receptor, Brad M. Binder, Ronan C. O'Malley, Wuyi Wang, Tobias C. Zutz, Anthony B. Bleeker Dec 2005

Ethylene Stimulates Nutations That Are Dependent On The Etr1 Receptor, Brad M. Binder, Ronan C. O'Malley, Wuyi Wang, Tobias C. Zutz, Anthony B. Bleeker

Brad M. Binder

Ethylene influences a number of processes in Arabidopsis (Arabidopsis thaliana) through the action of five receptors. In this study, we used high-resolution, time-lapse imaging to examine the long-term effects of ethylene on growing, etiolated Arabidopsis seedlings. These measurements revealed that ethylene stimulates nutations of the hypocotyls with an average delay in onset of over 6 h. The nutation response was constitutive in ctr1-2 mutants maintained in air, whereas ein2-1 mutants failed to nutate when treated with ethylene. Ethylene-stimulated nutations were also eliminated in etr1-7 loss-of-function mutants. Transformation of the etr1-7 mutant with a wild-type genomic ETR1 transgene rescued the nutation ...


Ethylene-Binding Activity, Gene-Expression Levels, And Receptor-System Output For Ethylene-Receptor Family Members From Arabidopsis And Tomato, Brad Binder, Ronan O'Malley, Fernando Rodriguez, Jeffrey Esch, Philip O'Donnell, Harry Klee, Anthony Bleecker Feb 2005

Ethylene-Binding Activity, Gene-Expression Levels, And Receptor-System Output For Ethylene-Receptor Family Members From Arabidopsis And Tomato, Brad Binder, Ronan O'Malley, Fernando Rodriguez, Jeffrey Esch, Philip O'Donnell, Harry Klee, Anthony Bleecker

Brad M. Binder

Ethylene signaling in plants is mediated by a family of ethylene receptors related to bacterial two-component regulators. Expression in yeast of ethylene-binding domains from the five receptor isoforms from Arabidopsis thaliana and five-receptor isoforms from tomato confirmed that all members of the family are capable of high-affinity ethylene-binding activity. All receptor isoforms displayed a similar level of ethylene binding on a per unit protein basis, while members of both subfamily I and subfamily II from Arabidopsis showed similar slow-release kinetics for ethylene. Quantification of receptor-isoform mRNA levels in receptor-deficient Arabidopsis lines indicated a direct correlation between total message level and ...


Short-Term Growth Responses To Ethylene In Arabidopsis Seedlings Are Ein3/Eil1 Independent, Brad Binder, Laura Mortimore, Anna Stepanova, Joseph Ecker, Anthony Bleecker Sep 2004

Short-Term Growth Responses To Ethylene In Arabidopsis Seedlings Are Ein3/Eil1 Independent, Brad Binder, Laura Mortimore, Anna Stepanova, Joseph Ecker, Anthony Bleecker

Brad M. Binder

Kinetic studies indicate there are two phases to growth inhibition by ethylene for the hypocotyls of etiolated Arabidopsis seedlings. Phase I is transient, while phase II results in sustained growth inhibition. The EIN2 membrane protein is required for both the first and second phases of growth inhibition by ethylene, while the transcription factors EIN3 and EIL1 are required for the second phase but not the first phase. The first phase lasts no more than 2 h. It is less sensitive to the ethylene response inhibitor 1-methylcyclopropene and more sensitive to ethylene than the second phase. The first phase shows adaptation ...


Arabidopsis Seedling Growth Response And Recovery To Ethylene. A Kinetic Analysis, Brad Binder, Ronan O'Malley, Wuyi Wang, Jeannette Moore, Brian Parks, Edgar Spalding, Anthony Bleecker Dec 2003

Arabidopsis Seedling Growth Response And Recovery To Ethylene. A Kinetic Analysis, Brad Binder, Ronan O'Malley, Wuyi Wang, Jeannette Moore, Brian Parks, Edgar Spalding, Anthony Bleecker

Brad M. Binder

Responses to the plant hormone ethylene are mediated by a family of five receptors in Arabidopsis that act in the absence of ethylene as negative regulators of response pathways. In this study, we examined the rapid kinetics of growth inhibition by ethylene and growth recovery after ethylene withdrawal in hypocotyls of etiolated seedlings of wild-type and ethylene receptor-deficient Arabidopsis lines. This analysis revealed that there are two phases to growth inhibition by ethylene in wild type: a rapid phase followed by a prolonged, slower phase. Full recovery of growth occurs approximately 90 min after ethylene removal. None of the receptor ...


A Model For Ethylene Receptor Function And 1-Methylcyclopropene Action, Brad Binder, A. Bleecker Dec 2002

A Model For Ethylene Receptor Function And 1-Methylcyclopropene Action, Brad Binder, A. Bleecker

Brad M. Binder

This paper provides a comprehensive review of current understanding of ethylene receptors, signal transduction using Arabidopsis as the model plant. It discusses the current theory on the mechanism of ethylene binding and proposes a mechanism for the action of ethylene response inhibitors such as 1-MCP.


A Copper Cofactor For The Ethylene Receptor Etr1 From Arabidopsis, Brad Binder, Fernando Rodríguez, Jeffrey Esch, Anne Hall, G. Schaller, Anthony Bleecker Jan 1999

A Copper Cofactor For The Ethylene Receptor Etr1 From Arabidopsis, Brad Binder, Fernando Rodríguez, Jeffrey Esch, Anne Hall, G. Schaller, Anthony Bleecker

Brad M. Binder

The ETR1 receptor from Arabidopsis binds the gaseous hormone ethylene. A copper ion associated with the ethylene-binding domain is required for high-affinity ethylene-binding activity. A missense mutation in the domain that renders the plant insensitive to ethylene eliminates both ethylene binding and the interaction of copper with the receptor. A sequence from the genome of the cyanobacterium Synechocystis sp. strain 6803 that shows homology to the ethylene-binding domain of ETR1 encodes a functional ethylene-binding protein. On the basis of sequence conservation between the Arabidopsis and the cyanobacterial ethylene-binding domains and on in vitro mutagenesis of ETR1, a structural model for ...