Presentation on theme: "Plant Hormone Signaling and the Ubiquitin-proteasome System Serena J. Gregory March 30 th, 2011."— Presentation transcript:
Plant Hormone Signaling and the Ubiquitin-proteasome System Serena J. Gregory March 30 th, 2011
Objectives Introduction Overview of ubiquitin and the Ub/proteasome pathway Hormone signaling – Auxin signaling – Ethylene signaling – Abscisic Acid signaling Summary
Growth and development in plants
Hormone signaling through protein destruction
The ubiquitin molecule 76 amino acid protein Contains a -grasp fold Di-glycine tail on the C-terminus Covalently attached to target proteins Vierstra, R.D. (2009)
Schematic overview of the ubiquitin pathway Vierstra, R.D. (2009)
The Ubiquitin Code Ye Y and Rape M. (2009).
Predicted number of genes that encode each UPS component in plants E1- 2 genes E2- 37 genes E3- >1400 Vierstra, R.D. (2009)
Target protein specificity is achieved through E3s Three types of E3s – RING – U-box – HECT Structure can be either single subunit or multi subunit Santner, A. & Estelle, M.(2010)
RING E3s RING (Really Interesting New Gene) 465 RING proteins 70 aa motif characterized as the ring finger Zinc-binding motif that binds to E2s Santner, A. & Estelle, M.(2010)Vierstra, R.D. (2009)
Multi subunit RING E3s Components – RING protein RBX1 (RING Box 1) – Cullin scaffold-like protein – Adaptor subunits – Additional substrate-recognition protein Types – SCF (Skp-Cullin-F-box) – CUL3-BTB (Broad-complex, Tramtrack, Bric-a-Brac) – CUL4-DDB1 (DNA-DAMAGE BINDING 1) Santner, A. & Estelle, M.(2010)
U-box E3s Modified RING-finger domain 64 U-Box proteins 70 aa U-box domain Does not use zinc ions to stabilize structure Santner, A. & Estelle, M.(2010)
HECT E3s Homology to E6-AP C Terminus (HECT) Smallest E3 subfamily 20 HECT proteins 350 aa motif Contains an Ub-binding and E2 binding site Santner, A. & Estelle, M.(2010)
SCF complex assembly and hormone signaling SCF-most common E3 Discovered through mutant screens Exposed to auxin or auxin transport inhibitors Disruptions of components of the SCF E3s CUL1 mutants were discovered that are auxin resistant Mark Estelle
Objectives Introduction Overview of ubiquitin and the Ub/proteasome pathway Hormone signaling – Auxin signaling – Ethylene signaling – Absicic Acid signaling Summary
Hormone signaling in plants Controls communication – Internally – Neighboring cells – Long distance between different organ systems Three step process – Signal perception, signal transduction and response Phytohormones – Produced by individual cells instead of gland secretion
Structures of Phytohormones
Functional Interactions of Phytohormones Jailais, Y. & Chory, J. (2010).
Plant hormone receptors Ethylene: ETR1 (histidine kinases) Cytokinins: CRE1 (histidine kinases) Auxin: TIR1 and IAA proteins GA: GID1 (regulates GID2 and DELLA) Jasmonic acid: COI1 and JAZs ABA: GTG1 and GTG2 BR: BRI1 (LRR-RLK)
Sites of plant hormone perception Santner, A. & Estelle, M. (2009).
Objectives Introduction Overview of the ubiquitin and the Ub/proteasome pathway Hormone signaling – Auxin signaling – Ethylene signaling – Absicic Acid signaling Summary
Auxin Signaling From the Greek auxein, meaning to grow. Described by Charles Darwin in 1880 as a growth promoting substance that moves in plants Responsible for – Cell division/elongation – Organogenesis – Prevents senescence Transported via influx transporter proteins and efflux transporter proteins Act primarily through Auxin Response Transcription Factors (ARFs)
Auxin signaling with ARFs
F-box protein TIR1 is the auxin receptor Vierstra, R.D. (2009)
Crystal structure of TIR1 Vierstra, R.D. (2009)
Jasmonic Acid and Gibberellic Acid Vierstra, R.D. (2009) F-box protein- CORONATE INSENSITIVE 1 (COI1) Target protein- JA-ZIM domain repressor proteins (JAZ) Transcription factor- MYB DOMAIN PROTEIN 2 (MYB2) F-box proteins- SLEEPY 1 AND SNEEZY 1 GA receptor- GA-INSENSITIVE DWARF 1 (GID1) Target protein- Della repressor proteins
Ethylene Signaling Gaseous hormone – Promote fruit ripening and leaf abscission and plant senescence – Resistance to pathogens and stress response Triple Response Phenotype – Dark grown seedlings exposed to ethylene – Very distinctive phenotype
The Ethylene Triple-Response Phenotype
Mutant testing ethylene insensitive WT
J. M. Alonso, J. R. Ecker 2001 AIRETHYLENE
Abscisic Acid Signaling Isoprenoid compound Responsible for – Control of seed dormancy – Drought response Controversial receptors – CHLH (GUN5 in Arabidopsis) – GCR2 Established receptors – GTG1 and GTG2
The RNA-binding protein FCA is an abscisic acid receptor Razem FARazem FA, El-Kereamy A, Abrams SR, Hill RD. Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, CanadaEl-Kereamy AAbrams SRHill RD Abstract The phytohormone abscisic acid (ABA) regulates various physiological processes in plants. The molecular mechanisms by which this is achieved are not fully understood. Genetic approaches have characterized several downstream components of ABA signaling, but a receptor for ABA has remained elusive. Although studies indicate that several ABA response genes encode RNA-binding or RNA-processing proteins, none has been found to be functional in binding ABA. Here we show that FCA, an RNA-binding protein involved in flowering, binds ABA with high affinity in an interaction that is stereospecific and follows receptor kinetics. The interaction between FCA and ABA has molecular effects on downstream events in the autonomous floral pathway and, consequently, on the ability of the plant to undergo transition to flowering. We further show that ABA binding exerts a direct control on the FCA-mediated processing of precursor messenger RNA. Our results indicate that FCA is an ABA receptor involved in RNA metabolism and in controlling flowering time.
FCA does not bind abscisic acid Risk JMRisk JM, Macknight RC, Day CL.Macknight RCDay CL Biochemistry Department, University of Otago, Dunedin 9054, New Zealand. Abstract The RNA-binding protein FCA promotes flowering in Arabidopsis. Razem et al. reported that FCA is also a receptor for the phytohormone abscisic acid (ABA). However, we find that FCA does not bind ABA, suggesting that the quality of the proteins assayed and the sensitivity of the ABA-binding assay have led Razem et al. to erroneous conclusions. Because similar assays have been used to characterize other ABA receptors, our results indicate that the ABA- binding properties of these proteins should be carefully re- evaluated and that alternative ABA receptors are likely to be discovered.
Sites of plant hormone perception Santner, A. & Estelle, M. (2009)
Abscisic Acid Signaling Two-fold process Blocks degradation of ABI5 preventing ubiquitination by KEG Promotes degradation of ABI3 by increasing AIP2 expression Vierstra, R.D. (2009)
Effects of abscisic acid signaling on stomata
Summary The ubiquitin-26S proteasome system plays a prominent regulatory role in plant hormone signaling E3 ubiquitin ligases represent a new class of hormone signaling receptors E3 activities can be manipulated by small molecules could lead to new pharmacological treatments Elucidation of all E3 ligases will potentially uncover other interesting signaling pathways and mechanisms
References Vierstra, R.D. (2009). The ubiquitin-26S proteasome system at the nexus of plant biology. Nat. Rev. Mol. Cell Biol. 10, 385–397. Ye Y and Rape M. (2009). Building ubiquitin chains: E2 enzymes at work. Nat. Rev. Mol. Cell Biol. 10(11): Santner, A. & Estelle, M. (2009). Recent advances and emerging trends in plant hormone signaling. Nature 459, 1071–1078. Santner, A. & Estelle, M.(2010). The ubiquitin-proteasome system regulates plant hormone signaling. The Plant Journal. 61, Jailais, Y. & Chory, J. (2010). Unraveling the paradoxes of plant hormone signaling integration. Nat. Structural & Mol. Biol. 17(6): Benavente, L. M. & Alonso, J. M. (2006). Molecular mechanisms of ethylene signaling in Arabidopsis. Mol. BioSyst. 2,
References J. M. Alonso, J. R. Ecker, The Ethylene Pathway: A Paradigm for Plant Hormone Signaling and Interaction. Science's STKE (2001), Tan, X. et al. (2007). Mechanism of auxin perception by the TIR1 ubiquitin ligase. Nature 446, 640–645. Pandey, S., Nelson, D. C. & Assmann, S. M. (2009). Two novel GPCR-type G proteins are abscisic acid receptors in Arabidopsis. Cell 136, 136–148. Christmann, Alexander & Grill, Erwin. (2009). Are GTGs ABA's biggest fans?. Cell, 136. Retrieved from biggest-fans/ htmlhttp://www.biomedsearch.com/nih/Are-GTGs-ABAs- biggest-fans/ html Risk J.M. Macknight R.C. (2008). FCA does not bind abscisic acid. Nature 456(7223): E5-6.