Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format
© Fujikawa et al.; licensee springer. 2014
Received: 9 September 2013
Accepted: 27 May 2014
Published: 28 June 2014
The rice interactome, in which a network of protein-protein interactions has been elucidated in rice, is a useful resource to identify functional modules of rice signal transduction pathways. Protein-protein interactions occur in cells in two ways, constitutive and regulative. While a yeast-based high-throughput method has been widely used to identify the constitutive interactions, a method to detect the regulated interactions is rarely developed for a large-scale analysis.
A split luciferase complementation assay was applied to detect the regulated interactions in rice. A transformation method of rice protoplasts in a 96-well plate was first established for a large-scale analysis. In addition, an antibody that specifically recognizes a carboxyl-terminal fragment of Renilla luciferase was newly developed. A pair of antibodies that recognize amino- and carboxyl- terminal fragments of Renilla luciferase, respectively, was then used to monitor quality and quantity of interacting recombinant-proteins accumulated in the cells. For a proof-of-concept, the method was applied to detect the gibberellin-dependent interaction between GIBBERELLIN INSENSITIVE DWARF1 and SLENDER RICE 1.
A method to detect regulated protein-protein interactions was developed towards establishment of the rice interactome.
Interactome analysis collects data of protein-protein interactions occurring in cells. A useful application of the analysis is to identify functional modules of signal transduction pathways in which a network of protein-protein interactions mediate the signals (Barabasi et al. ; Pawson and Nash ). Protein-protein interactions occur in cells in two ways, constitutive and regulative. Constitutive protein interactions occur all the time in cells. In many cases, even when interaction is examined in vitro, a constitutive interaction is detectable. On the other hand, regulated protein interactions occur when cells are exposed to selected environments such as stress and environmental cue. When cells are exposed to selected environments, some of proteins change their subcellular localization to interact with other proteins. Alternatively, cells may produce small compound (i.e., hormone) that mediates a protein-protein interaction (Ritter and Hall ), or a protein in the cells be modified (i.e., phosphorylation) to attract other proteins (Pawson ). Importantly, regulated protein-protein interactions are often at pivotal positions within the interaction network in a signal transduction pathway (Wehr et al. ).
Because plants highly adaptive to different environments, signal transduction pathways in plants are thought to be complicated (Vanstraelen and Benkova ; Santner and Estelle ). Genomes of Oryza genus have been modified to adapt diverse environments and increase the value as a crop (Vaughan et al. ). Hence, genetic analysis of Oryza sativa, rice, is recently spurred considerable interest to understand the genomic evolution. However, rice interactome analysis, which would reveal the molecular mechanism of the rice adaptation, has been limited to constitutive protein-protein interactions that are determined by the yeast two-hybrid assay (Gu et al. ; Seo et al. ). Although the assay would detect protein-protein interactions in absent or present of certain regulators, it is difficult to detect regulated protein-protein interactions that may occur in cellular conditions specific to rice.
The split luciferase complementation (SLC) assay is one of methods that detect regulated protein-protein interactions in situ (Ozawa et al. ; Kaihara et al. ; Luker et al. ). In the assay, N- and C-terminal fragments of luciferase are genetically fused to a protein pair of interest. The luciferase activity, which emits light through oxidation of a substrate, is complemented when the protein pair interacts with each other, but ceases when the protein pair does not interact. Hence, one can determine a protein interaction by a flash of light. Because the assay is capable of identifying not only interaction but also dissociation of protein pairs, it is suitable for analyzing kinetics of protein interactions in situ (Luker et al. ). Moreover, the result of the SLC assay is reproducible with high signal-to-background ratios, which allows for conducting interactome analysis, at least, in animal cells (Cassonnet et al. ).
In plants, the assay was first applied to detect a histone-histone interaction in protoplasts of Arabidopsis leaves (Fujikawa and Kato ). Since then, the assay has been applied to detect interactions between membrane proteins (Kato et al. ), bacterial effector proteins and their protein targets (Chen et al. ), auxin response factors (Li et al. ), 14-3-3 regulator proteins (Gehl et al. ), coiled-coil–nucleotide-binding site–leucine-rich repeat (CC-NB-LRR) protein (Inoue et al. ), and kinase (Schmidt et al. ). We previously showed with the SLC assay that protein-protein interactions in Arabidopsis protoplasts are sensitive to environmental conditions (Kato and Bai ), indicating that the SLC assay would be well-suited for analyzing regulated protein-protein interactions in plant cells.
Currently, however, the use of the SLC in plants is limited to detecting constitutive protein-protein interactions and only in dicot model plants such as Arabdidopsis and tobacco, but not in monocot plants such as rice. Comparative transcriptional analyses revealed that rice and Arabidopsis differently transduce a stress signal (Yazaki et al. ; Narsai and Whelan ). Moreover, gene annotation and transcriptome analyses revealed that rice and Arabidopsis differently regulate the flow of potassium in the plasma membrane as a result of stress and environmental cue (Very et al. ; Ma et al. ). Potassium is the major inorganic cation in the cytoplasm, and known to regulate the activities of many enzymes including ones involved in signal transductions in plant cells (Wang and Wu ). These analyses support the idea that rice may possess a unique network in regulated interactions, which could not be identified when a protein-interaction assay is conducted in different organisms (i.e. Arabidopsis and yeast).
Here we present a Renilla luciferase-based technology breakthrough that allows the identification of regulated protein-protein interactions in rice in near-real time, which can also be expanded to a large-scale analysis.
Results and discussion
A transformation method for rice protoplasts in a 96-well plate was established
An antibody that recognizes the C- terminal fragment of Renilla luciferase was generated
Luminescence emitted from protoplasts depends on not only the affinity (dissociation constant) of a protein pair that is fused to NRLuc and CRLuc, respectively, but also the amounts the recombinant proteins accumulated in the protoplasts (Kato et al. ). Because luminescence units determined in the assay are relative but not absolute, it remains a challenge to deduce an absolute value of the dissociation constant. However, the anti-CRLuc antibody developed in this study will allows addressing the question whether low luminescence in the assay is caused by a low affinity of the protein pair or low levels of the protein accumulation in one or both of the protein pair as discussed in the following sections.
Histone2A-histone2B interaction was detected with a high signal-to-background ratio
To examine whether a protein interaction was detected by bioluminescence in rice protoplasts, relative luminescence units (RLU) were measured soon after adding the luciferase substrate to the culture solution in each well of the 96-well plate but before lysing the protoplasts for the immunoblot assay. Because H2A interacts with H2B (Fujikawa and Kato ), we expected rice protoplasts expressing NRLuc-H2A and CRLuc-H2B to emit luminescence light at high levels upon addition of viviRen®, the substrate for Renilla luciferase. On the other hand, we expected rice protoplasts expressing NRLuc-H2A and CRLuc-SLR1 or NRLuc-GID1 and CRLuc-H2B to emit luminescence light at very low levels, similar to mock-transformed protoplasts, because these protein pairs would not interact with each other. The rice protoplasts expressing NRLuc-H2A and CRLuc-H2B showed 21-fold higher RLU, compared to the mock transformed protoplasts (Figure 2B). On the other hand, the protoplasts expressing NRLuc-H2A and CRLuc-SLR1 or those expressing NRLuc-GID1 and CRLuc-SRL1 showed RLU at low levels relative to the mock transformed protoplasts. This suggested that rice protoplasts are suitable to determine protein-protein interactions by the SLC assay in a large-scale format as we previously found in Arabidopsis protoplasts (Fujikawa and Kato ).
Gibberellin-dependent interaction of GID1 and SLR1 was detected in near-real time
Adaptation of plant growth and development to the environment is largely achieved through hormone production and distribution in the plant (Vanstraelen and Benkova ; Santner and Estelle ). Hence, signal transduction pathways and a protein interaction network in plant cells are expected to be modulated by hormones. To test whether our system could detect hormone-dependent protein interactions, we took advantage of the previously characterized interaction between two components of gibberellin signaling, GID1 and SLR1 (Ueguchi-Tanaka et al. ).
GAs are a large family of plant hormones that widely play roles in plant development, such as seed germination, stem elongation, leaf expansion, flowering, and pollen maturation (Thomas and Sun ). A breakthrough in the GA signaling study is the discovery of a soluble GA receptor, GID1 (Ueguchi-Tanaka et al. ). The gid1 mutant rice shows dwarf and insensitive to the exposure of exogenous GA. Structural analyses of GID1 proteins of rice and Arabidopsis revealed that an N-terminal portion of the protein functions as a lid to bind and cover GA (Murase et al. ; Shimada et al. ). The N-terminal lid is also found to involve in the GA-dependent interaction with SLR1, the repressor in GA signaling (Murase et al. ; Shimada et al. ). Interestingly for us, while the rice (Oryza sativa) genome encodes one GID1 protein OsGID1, the Arabidopsis (Arabidopsis thaliana) genome encodes the OsGID1 orthologous protein AtGID1a and the paralogous proteins AtGID1b (Nakajima et al. ). AtGID1b constitutively interacts with SLR1 without GA (Yamamoto et al. ). This supports the idea that rice and Arabidopsis cells may have different networks in the GA-regulated protein-interactions.
Interaction of GID1 and SLR1 was not affected by other plant hormones in rice protoplasts
Although other hormones increased RLU, the increases were not significant (P > 0.10) compared to the mock transformed protoplasts. We also confirmed by immunoblot assays that amounts of the recombinant proteins accumulated in the transformed protoplasts were not affected by different plant hormones (Figure 4B). This suggested that the GID1-SLR1 interaction was mediated by gibberellin specifically in the rice cells. This also showed that the SLC assay is capable of detecting a protein-protein interaction that is regulated by a hormone.
We previously discussed about advantage and disadvantage of SLC by comparing the results of SLC in Arabidopsis protoplasts with that of other interaction assays such as FRET (Fluorescence resonance energy transfer) and Co-IP (Co-immunoprecipitaiton) assay (Fujikawa and Kato ; Kato et al. ). We, furthermore, used the SLC assay to detect a light-dependent interaction in Arabidopsis protoplasts (Kato et al. ). Here we showed that the SLC is capable of detecting a protein-protein interaction that is regulated by a hormone in rice protoplasts in near-real time. Because pDuEx vectors used in this SLC assay contain the Gateway® recombinant sites (Fujikawa and Kato ), one can relatively easily construct a large number of expression vectors. Moreover, four different types of pDuEx vectors that allow fusing NRLuc or CRLuc to N- or C-terminal ends of a target protein, were constructed (GenBank: EF565883, EF56588, GU370778 and GU370779). All together, these provide a suite of vectors for a large-scale SLC assay in rice protoplasts. The suite would aid in revealing regulated protein-protein interactions in rice and other monocot crop cells in a large-scale format.
The coding sequences in the cDNAs including a stop codon (GID1; GIBBERELLIN INSENSITIVE DWARF1, Genbank Acc. No. AB211399, SLR1; SLENDER RICE1 Genbank Acc. No. AB262980) were amplified by PCR using pGBKT7-GID1 and pGADT7-SLR1 (Ueguchi-Tanaka et al. ) as the templates, respectively. The products of PCR were then cloned into a pDONR/Zeo vector (Invitrogen, CA). The resultant plasmids were subsequently used to clone into pDuExAn6 or D7 expression vectors (Fujikawa and Kato ) with the Gateway® cloning system (Invitrogen, CA).
Rice cultivation and treatments
Mature seeds of rice (Oryza sativa L. Japonica) were soaked in water for 3 days at 28°C in the dark after being sterilized with 50% (v/v) bleach for 10 min. The seeds were then placed on soil in a pot (8 cm wide by 7.5 cm high) with Kimura B solution [365 μM (NH4)2SO4, 547 μM MgSO4, 183 μM KNO3, 365 μM Ca(NO3)2, 182 μM KH2PO4, 19 μM Fe-EDTA, 48.7 μM H3BO3, 9 μM MnSO4, 0.3 μM CuSO4, 0.7 μM ZnSO4, 0.099 μM Na2MoO4] containing 1 nM uniconazole. The seedlings were incubated at 28°C in 16 h of light (light intensity; 100 μmol m−2 sec−1) and 8 h of dark for 1 to 2 weeks.
Protoplast preparation and transformation
Rice shoots (0.5 - 1.0 g, 1–2 week old) were cut into 2 mm-wide sections toward the leaf sheath by using a surgical knife (No.21 FETHER, Osaka, Japan). Protoplasts were isolated by enzymatic hydrolysis using cellularase RS (Yakult pharmaceutical industry. Tokyo, Japan) and macerozyme R10 (Yakult pharmaceutical industry). The pieces of the rice shoots were first incubated with the enzyme solution (4% cellulase RS and 2% macerozyme R10, 0.6 M mannitol, 20 mM MES pH 5.7, 20 mM KCl, and 10 mM CaCl2) for 4 h on a rotary platform shaker at 35 rpm/min after vacuuming the samples for 30 min. The samples were then filtered through a 40 μm nylon mesh to remove clumps. The isolated protoplasts were washed two times with 10 ml of W5 buffer (0.125 M CaCl2, 5 mM KCl, and 2 mM MES pH 5.7) by centrifugation at 200 × g for 3 min. The protoplasts were further suspended in 10 ml of W5 buffer and placed at 4°C for 30 min. The protoplasts were collected by centrifugation at 200 × g for 3 min, and re-suspended in MMg solution (0.4 M mannitol, 15 mM MgCl2, and 4 mM MES pH 5.7) so that the concentration of the protoplasts in the solution would be 4 × 105 cells/ml. Protoplast transformation was conducted using a 96-well plate (U-bottom plate; U96 MicroWell Plates, Nalge Nunc International, NY) based on the method previously published (Kato and Jones ). Briefly, the expression vectors dissolved in 10 μl of distilled water were used to transform 1.6 × 104 protoplasts in 40 μl of the MMg buffer. After adding 60 μl of polyethylene glycol solution containing 40% (w/v) PEG4000 (Sigma-Fluka, MO), 0.2 M mannitol and 0.1 M CaCl2 to the wells, the plate was vortexed with a digital vortex mixer (GENIE2, Scientific industries) at 900 rpm for 15 sec. The transformed protoplasts in each were then washed with 200 μl of W5 buffer. To replace the solution in each well, the plate was centrifuged at 200× g for 3 min at room temperature and 200 μl of the supernatant was removed from each well. The remaining protoplasts were washed four-times in 200 μl of W5 buffer. The transformed protoplasts (1.6 × 104 cells) were suspended in 100 μl of W5 buffer. Finally, the plate was shaken for 5 sec and incubated in the dark at 28°C overnight.
Measuring luciferase luminescence
Luminescence was measured by a microplate luminometer (ARVOx4 2030 Multilabel Reader, Perkin Elmer, MA) immediately after adding 10 μl of 0.12 mM ViviRen® (Promega) in each well where the transformed protoplasts were incubated in the 100 μl solution. The luminescence signals in each well were integrated for 0.5 sec in one measurement every 1.5 min. When hormones were added, luminescence signals in a 96-well plate were measured about every 6 min for 90 min or 120 min before and after adding plant hormones. GA3 and GA4 were purchased from Sigma-Fluka. Abscisic acid (ABA) and jasmonic acid (JA) were from Wako Pure Chemical (Osaka, Japan). Indoleacetic acid (IAA), kinetin (KT) and salicylic acid (SA) were from Nakarai tesque (Kyoto Japan).
Generation of antibody
cDNA encoding the C-terminal fragment of Renilla luciferase was amplified from pDUExD7 (Fujikawa and Kato ) by PCR using CRLuc-F (5’-CCGGAATTCAAGCCCGACGTCCAGATT-3’) and CRLuc-R (5’-CCGCTCGAGCTGCTCGTTCTTCAGCACGCG-3’) primers. The resulted PCR product was subcloned into pGEX5X-1 (GE Healthcare, WI). The resultant plasmid was introduced into BL21, and the GST-fused CRLuc protein fragment was induced by adding 0.1 mM IPTG for 12 hr at 30°C. The fusion protein was purified by glutathione Sepharose 4B according to the manufacture’s instruction (GE Helthcare). The purified protein was used to obtain a polyclonal antibody using rabbit.
Rice protoplasts (3.2 × 104 cells) suspended in 10 μl of W5 buffer were homogenized with 10 μl of 2x SDS-PAGE sample buffer containing 0.2 M Tris–HCl (pH 6.8), 20% (v/v) glycerol, 4% (w/v) SDS, 0.15 M sodium chloride, 2% (v/v) Triton X-100, 0.005% (w/v) brome phenol blue, 4% (v/v) 2-mercaptoethanol, and 6 M urea. The samples were boiled for 5 min and loaded on a 5-20% gradient polyacrylamide gel (E-R520L, Atto Corp., Tokyo, Japan). Proteins in the gel were then transferred to a polyvinylidene difluoride membrane (Immobilon-P transfer membrane, Millipore Corp., MA) using a semidry electrotransfer (15 V constant). The membrane was incubated for 90 min in a blocking solution [4% (w/v) nonfat dry milk and 0.1% (v/v) Triton X-100 in 10 mM Tris–HCl, pH 7.4, 150 mM sodium chloride), and further incubated overnight with mouse monoclonal anti NRLuc (1:20,000, MAB4400, Millipore Corp) or anti-CRLuc rabbit serum (1:10,000) in the blocking solution. The binding of these antibodies was detected with an ECL Western blotting system (Amersham Bioscience/GE Healthcare, WI).
FY was a postdoctoral associate in LSU at the early stage of the study.
The study was supported by the USDA Cooperative State Research, Education and Extension Service – National Research Initiative – Plant Genome Program, award no. 2006-35604-16627 for NK. We thank Dr. Aaron Smith for his comments and suggestions on the manuscript.
- Barabasi AL, Gulbahce N, Loscalzo J: Network medicine: a network-based approach to human disease. Nat Rev Genet 2011, 12(1):56–68. doi:10.1038/nrg2918 doi:10.1038/nrg2918 10.1038/nrg2918PubMed CentralView ArticlePubMedGoogle Scholar
- Bart R, Chern M, Park C-J, Bartley L, Ronald P: A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts. Plant Methods 2006, 2(1):13. 10.1186/1746-4811-2-13PubMed CentralView ArticlePubMedGoogle Scholar
- Cassonnet P, Rolloy C, Neveu G, Vidalain PO, Chantier T, Pellet J, Jones L, Muller M, Demeret C, Gaud G, Vuillier F, Lotteau V, Tangy F, Favre M, Jacob Y: Benchmarking a luciferase complementation assay for detecting protein complexes. Nat Methods 2011, 8(12):990–992. doi:10.1038/nmeth.1773 doi:10.1038/nmeth.1773 10.1038/nmeth.1773View ArticlePubMedGoogle Scholar
- Chen H, Zou Y, Shang Y, Lin H, Wang Y, Cai R, Tang X, Zhou JM: Firefly luciferase complementation imaging assay for protein-protein interactions in plants. Plant physiology 2008, 146(2):368–376. doi:10.1104/pp.107.111740 doi:10.1104/pp.107.111740 10.1104/pp.107.111740PubMed CentralView ArticlePubMedGoogle Scholar
- Datta K, Datta S: Transformation of Rice via PEG-Mediated DNA Uptake into Protoplasts. In Plant Cell Culture Protocols, vol 111. Methods In Molecular Biology‚Ñ¢. Edited by: Hall R. Humana Press, Humana Press; 1999:335–347. doi:10.1385/1–59259–583–9:335 doi:10.1385/1-59259-583-9:335 10.1385/1-59259-583-9:335View ArticleGoogle Scholar
- Fujikawa Y, Kato N: Split luciferase complementation assay to study protein-protein interactions in Arabidopsis protoplasts. Plant J 2007, 52(1):185–195. 10.1111/j.1365-313X.2007.03214.xView ArticlePubMedGoogle Scholar
- Gehl C, Kaufholdt D, Hamisch D, Bikker R, Kudla J, Mendel RR, Hansch R: Quantitative analysis of dynamic protein-protein interactions in planta by a floated-leaf luciferase complementation imaging (FLuCI) assay using binary Gateway vectors. The Plant journal 2011, 67(3):542–553. doi:10.1111/j.1365–313X.2011.04607.x doi:10.1111/j.1365-313X.2011.04607.x 10.1111/j.1365-313X.2011.04607.xView ArticlePubMedGoogle Scholar
- Gu H, Zhu P, Jiao Y, Meng Y, Chen M: PRIN: a predicted rice interactome network. BMC Bioinformatics 2011, 12: 161. doi:10.1186/1471–2105–12–161 doi:10.1186/1471-2105-12-161 10.1186/1471-2105-12-161PubMed CentralView ArticlePubMedGoogle Scholar
- Inoue H, Hayashi N, Matsushita A, Xinqiong L, Nakayama A, Sugano S, Jiang CJ, Takatsuji H: Blast resistance of CC-NB-LRR protein Pb1 is mediated by WRKY45 through protein-protein interaction. Proc Natl Acad Sci U S A 2013, 110(23):9577–9582. doi:10.1073/pnas.1222155110 doi:10.1073/pnas.1222155110 10.1073/pnas.1222155110PubMed CentralView ArticlePubMedGoogle Scholar
- Kaihara A, Kawai Y, Sato M, Ozawa T, Umezawa Y: Locating a protein-protein interaction in living cells via split Renilla luciferase complementation. Anal Chem 2003, 75(16):4176–4181. 10.1021/ac0300800View ArticlePubMedGoogle Scholar
- Kato N, Bai H (2010) Expression, localization, and interaction of SNARE proteins in Arabidopsis are selectively altered by the dark. Plant Signaling & Behavior 5(11)Google Scholar
- Kato N, Jones J: Split luciferase complementation assay. In Methods in Plant Development. Edited by: Henning L, Kohler C. Methods in Molecular Biology. Humana Press Inc, NY; 2010:359–376. 10.1007/978-1-60761-765-5_24Google Scholar
- Kato N, Fujikawa Y, Fuselier T, Adamou-Dodo R, Nishimura A, Sato MH (2009) Luminescence detection of SNARE-SNARE interaction in Arabidopsis protoplasts. Plant Mol Biol:Publsihed online 12(December 2009)Google Scholar
- Li JF, Bush J, Xiong Y, Li L, McCormack M: Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation. PLoS One 2011, 6(11):e27364. doi:10.1371/journal.pone.0027364 doi:10.1371/journal.pone.0027364 10.1371/journal.pone.0027364PubMed CentralView ArticlePubMedGoogle Scholar
- Luker KE, Smith MC, Luker GD, Gammon ST, Piwnica-Worms H, Piwnica-Worms D: Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals. Proc Natl Acad Sci U S A 2004, 101(33):12288–12293. doi:10.1073/pnas.0404041101 0404041101 [pii] doi:10.1073/pnas.0404041101 0404041101 [pii] 10.1073/pnas.0404041101PubMed CentralView ArticlePubMedGoogle Scholar
- Ma TL, Wu WH, Wang Y: Transcriptome analysis of rice root responses to potassium deficiency. BMC Plant Biol 2012, 12: 161. doi:10.1186/1471–2229–12–161 doi:10.1186/1471-2229-12-161 10.1186/1471-2229-12-161PubMed CentralView ArticlePubMedGoogle Scholar
- Murase K, Hirano Y, Sun TP, Hakoshima T: Gibberellin-induced DELLA recognition by the gibberellin receptor GID1. Nature 2008, 456(7221):459–463. doi:10.1038/nature07519 doi:10.1038/nature07519 10.1038/nature07519View ArticlePubMedGoogle Scholar
- Nakajima M, Shimada A, Takashi Y, Kim YC, Park SH, Ueguchi-Tanaka M, Suzuki H, Katoh E, Iuchi S, Kobayashi M, Maeda T, Matsuoka M, Yamaguchi I: Identification and characterization of Arabidopsis gibberellin receptors. Plant J 2006, 46(5):880–889. 10.1111/j.1365-313X.2006.02748.xView ArticlePubMedGoogle Scholar
- Narsai R, Whelan J: How unique is the low oxygen response? An analysis of the anaerobic response during germination and comparison with abiotic stress in rice and Arabidopsis. Front Plant Sci 2013, 4: 349. doi:10.3389/fpls.2013.00349 doi:10.3389/fpls.2013.00349 10.3389/fpls.2013.00349PubMed CentralView ArticlePubMedGoogle Scholar
- Ozawa T, Kaihara A, Sato M, Tachihara K, Umezawa Y: Split luciferase as an optical probe for detecting protein-protein interactions in mammalian cells based on protein splicing. Anal Chem 2001, 73(11):2516–2521. 10.1021/ac0013296View ArticlePubMedGoogle Scholar
- Pawson T: Specificity in signal transduction: from phosphotyrosine-SH2 domain interactions to complex cellular systems. Cell 2004, 116(2):191–203. 10.1016/S0092-8674(03)01077-8View ArticlePubMedGoogle Scholar
- Pawson T, Nash P: Protein-protein interactions define specificity in signal transduction. Genes & development 2000, 14(9):1027–1047.Google Scholar
- Ritter SL, Hall RA: Fine-tuning of GPCR activity by receptor-interacting proteins. Nat Rev Mol Cell Biol 2009, 10(12):819–830. doi:10.1038/nrm2803 doi:10.1038/nrm2803 10.1038/nrm2803PubMed CentralView ArticlePubMedGoogle Scholar
- Santner A, Estelle M: Recent advances and emerging trends in plant hormone signalling. Nature 2009, 459(7250):1071–1078. doi:10.1038/nature08122 doi:10.1038/nature08122 10.1038/nature08122View ArticlePubMedGoogle Scholar
- Schmidt R, Mieulet D, Hubberten HM, Obata T, Hoefgen R, Fernie AR, Fisahn J, San Segundo B, Guiderdoni E, Schippers JH, Mueller-Roeber B (2013) Salt-responsive ERF1 Regulates Reactive Oxygen Species-Dependent Signaling during the Initial Response to Salt Stress in Rice. The Plant cell, doi:10.1105/tpc.113.113068Google Scholar
- Seo YS, Chern M, Bartley LE, Han M, Jung KH, Lee I, Walia H, Richter T, Xu X, Cao P, Bai W, Ramanan R, Amonpant F, Arul L, Canlas PE, Ruan R, Park CJ, Chen X, Hwang S, Jeon JS, Ronald PC: Towards establishment of a rice stress response interactome. PLoS Genet 2011, 7(4):e1002020. doi:10.1371/journal.pgen.1002020 doi:10.1371/journal.pgen.1002020 10.1371/journal.pgen.1002020PubMed CentralView ArticlePubMedGoogle Scholar
- Shimada A, Ueguchi-Tanaka M, Nakatsu T, Nakajima M, Naoe Y, Ohmiya H, Kato H, Matsuoka M: Structural basis for gibberellin recognition by its receptor GID1. Nature 2008, 456(7221):520–523. doi:10.1038/nature07546 doi:10.1038/nature07546 10.1038/nature07546View ArticlePubMedGoogle Scholar
- Thomas SG, Sun TP: Update on gibberellin signaling. A tale of the tall and the short. Plant physiology 2004, 135(2):668–676. doi:10.1104/pp.104.040279 doi:10.1104/pp.104.040279 10.1104/pp.104.040279PubMed CentralView ArticlePubMedGoogle Scholar
- Ueguchi-Tanaka M, Ashikari M, Nakajima M, Itoh H, Katoh E, Kobayashi M, Chow TY, Hsing YI, Kitano H, Yamaguchi I, Matsuoka M: Gibberellin insensitive DWARF1 encodes a soluble receptor for gibberellin. Nature 2005, 437(7059):693–698. doi:10.1038/nature04028 doi:10.1038/nature04028 10.1038/nature04028View ArticlePubMedGoogle Scholar
- Ueguchi-Tanaka M, Nakajima M, Katoh E, Ohmiya H, Asano K, Saji S, Hongyu X, Ashikari M, Kitano H, Yamaguchi I, Matsuoka M: Molecular interactions of a soluble gibberellin receptor, GID1, with a rice DELLA protein, SLR1, and gibberellin. Plant Cell 2007, 19(7):2140–2155. 10.1105/tpc.106.043729PubMed CentralView ArticlePubMedGoogle Scholar
- Ueguchi-Tanaka M, Nakajima M, Motoyuki A, Matsuoka M: Gibberellin receptor and its role in gibberellin signaling in plants. Annu Rev Plant Biol 2007, 58: 183–198. 10.1146/annurev.arplant.58.032806.103830View ArticlePubMedGoogle Scholar
- Vanstraelen M, Benkova E: Hormonal interactions in the regulation of plant development. Annu Rev Cell Dev Biol 2012, 28: 463–487. doi:10.1146/annurev-cellbio-101011–155741 doi:10.1146/annurev-cellbio-101011-155741 10.1146/annurev-cellbio-101011-155741View ArticlePubMedGoogle Scholar
- Vaughan DA, Morishima H, Kadowaki K: Diversity in the Oryza genus. Curr Opin Plant Biol 2003, 6(2):139–146. 10.1016/S1369-5266(03)00009-8View ArticlePubMedGoogle Scholar
- Very AA, Nieves-Cordones M, Daly M, Khan I, Fizames C, Sentenac H (2014) Molecular biology of K transport across the plant cell membrane: What do we learn from comparison between plant species? J Plant Physiol, doi:10.1016/j.jplph.2014.01.011PubMedGoogle Scholar
- Wang Y, Wu WH: Potassium transport and signaling in higher plants. Annu Rev Plant Biol 2013, 64: 451–476. doi:10.1146/annurev-arplant-050312–120153 doi:10.1146/annurev-arplant-050312-120153 10.1146/annurev-arplant-050312-120153View ArticlePubMedGoogle Scholar
- Wehr MC, Reinecke L, Botvinnik A, Rossner MJ: Analysis of transient phosphorylation-dependent protein-protein interactions in living mammalian cells using split-TEV. BMC Biotechnol 2008, 8: 55. doi:10.1186/1472–6750–8-55 doi:10.1186/1472-6750-8-55 10.1186/1472-6750-8-55PubMed CentralView ArticlePubMedGoogle Scholar
- Weiss D, Ori N: Mechanisms of cross talk between gibberellin and other hormones. Plant physiology 2007, 144(3):1240–1246. doi:10.1104/pp.107.100370 doi:10.1104/pp.107.100370 10.1104/pp.107.100370PubMed CentralView ArticlePubMedGoogle Scholar
- Yamamoto Y, Hirai T, Yamamoto E, Kawamura M, Sato T, Kitano H, Matsuoka M, Ueguchi-Tanaka M: A rice gid1 suppressor mutant reveals that gibberellin is not always required for interaction between its receptor, GID1, and DELLA proteins. The Plant cell 2010, 22(11):3589–3602. doi:10.1105/tpc.110.074542 doi:10.1105/tpc.110.074542 10.1105/tpc.110.074542PubMed CentralView ArticlePubMedGoogle Scholar
- Yazaki J, Shimatani Z, Hashimoto A, Nagata Y, Fujii F, Kojima K, Suzuki K, Taya T, Tonouchi M, Nelson C, Nakagawa A, Otomo Y, Murakami K, Matsubara K, Kawai J, Carninci P, Hayashizaki Y, Kikuchi S: Transcriptional profiling of genes responsive to abscisic acid and gibberellin in rice: phenotyping and comparative analysis between rice and Arabidopsis. Physiol Genomics 2004, 17(2):87–100. doi:10.1152/physiolgenomics.00201.2003 doi:10.1152/physiolgenomics.00201.2003 10.1152/physiolgenomics.00201.2003View ArticlePubMedGoogle Scholar
- Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D, Wang J, Wang H: A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 2011, 7(1):30. 10.1186/1746-4811-7-30PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproductionin any medium, provided the original work is properly credited.