CHRK1, a chitinase-related receptor-like kinase in tobacco

A cDNA encoding a chitinase-related receptor-like kinase, designated CHRK1, was isolated from tobacco (Nicotiana tabacum). The C-terminal kinase domain (KD) of CHRK1 contained all of the conserved amino acids of serine/threonine protein kinases. The putative extracellular domain was closely related to the class V chitinase of tobacco and to microbial chitinases. CHRK1 mRNA accumulation was strongly stimulated by infection with fungal pathogen and tobacco mosaic virus. Amino acid-sequence analysis revealed that the chitinase-like domain of CHRK1 lacked the essential glutamic acid residue required for chitinase activity. The recombinant chitinase-like domain did not show any catalytic activity for either oligomeric or polymeric chitin substrates. The recombinant KD of CHRK1 exhibited autophosphorylation, but the mutant KD with a mutation in the essential ATP-binding site did not, suggesting that CHRK1 encoded a functional kinase. CHRK1 was detected in membrane fractions of tobacco BY2 cells. Furthermore, CHRK1-GFP fusion protein was localized in plasma membranes when it was expressed in animal cells. This is the first report of a new type of receptor-like kinase containing a chitinase-like sequence in the putative extracellular domain.     

CHRK1, a chitinase-related receptor-like kinase,plays a role in plant development and cytokinin homeostasis in tobacco

CHRK1 encodes a receptor-like kinase that contains a chitinase-related sequence in the extracellular domain in Nicotiana tabacum. In this study, we showed that CHRK1 is mainly expressed in the shoot apex region including leaf primordia and young leaves, and germinating seedlings and vascular tissues, based on GUS activity of transgenic tobacco plants carrying the CHRK1 promoter-GUS fusion gene. Transgenic tobacco plants in which CHRK1 expression was suppressed exhibited pleiotrophic developmental abnormality, including formation of proliferating shooty calli from emerging seedlings and severely altered seedling development. At the cellular level, ectopic cell proliferation, reduced cell specificity, and aberrant chloroplast development were observed. The transgenic lines contained 3-fold higher level of cytokinin than the wild-type plants. Consistently, the transgenic seedlings exhibited a typical cytokinin response in the absence of hormone, such as deetiolation under the dark. Based on these results, we propose that CHRK1 is involved in a developmental signaling pathway regulating cell proliferation/differentiation and the endogenous cytokinin levels in tobacco.     

The shooty callus induced by suppression of tobacco CHRK1 receptor-like kinase is a phenocopy of the tobacco genetic tumor

CHRK1 encodes a tobacco receptor-like kinase that contains a chitinase-like sequence in the extracellular domain. In a previous study, CHRK1-suppressed transgenic tobacco plants exhibited pleiotropic developmental abnormalities including spontaneous growth of shooty callus from emerging embryos in the absence of any exogenous hormones. In this study, we show that the CHRK1 shooty callus mimics tobacco genetic tumors in its morphology, physiology, and gene expression profiles. Similar to CHRK1 shooty callus, tobacco genetic tumors exhibit shooty callus morphology and hormone-independent shoot organogenesis. Both the CHRK1 callus and genetic tumors constitutively expressed KNOTTED1-type homeobox genes at the high levels, consistent with their vigorous shoot formation. These two types of calli exhibited cell death phenotypes, accompanied by high H₂O₂ production, increased ion leakage, and callose accumulation. Consistently, both types of calli constitutively expressed high levels of defense genes induced during pathogen-mediated HR cell death. These results, together with previous reports that both the CHRK1 shooty callus and tobacco genetic tumor contained high levels of cytokinin, indicate that CHRK1 shooty callus is a phenocopy of tobacco genetic tumor. CHRK1-mediated signal transduction may play a role in the formation of the genetic tumor in tobacco.     

Two LysM receptor molecules, CEBiP and OsCERK1, cooperatively regulate chitin elicitor signaling in rice

Chitin is a major molecular pattern for various fungi, and its fragments, chitin oligosaccharides, are known to induce various defense responses in plant cells. A plasma membrane glycoprotein, CEBiP (chitin elicitor binding protein) and a receptor kinase, CERK1 (chitin elicitor receptor kinase) (also known as LysM-RLK1), were identified as critical components for chitin signaling in rice and Arabidopsis, respectively. However, it is not known whether each plant species requires both of these two types of molecules for chitin signaling, nor the relationships between these molecules in membrane signaling. We report here that rice cells require a LysM receptor-like kinase, OsCERK1, in addition to CEBiP, for chitin signaling. Knockdown of OsCERK1 resulted in marked suppression of the defense responses induced by chitin oligosaccharides, indicating that OsCERK1 is essential for chitin signaling in rice. The results of a yeast two-hybrid assay indicated that both CEBiP and OsCERK1 have the potential to form hetero- or homo-oligomers. Immunoprecipitation using a membrane preparation from rice cells treated with chitin oligosaccharides suggested the ligand-induced formation of a receptor complex containing both CEBiP and OsCERK1. Blue native PAGE and chemical cross-linking experiments also suggested that a major portion of CEBiP exists as homo-oligomers even in the absence of chitin oligosaccharides.     

ROP GTPase-mediated auxin signaling regulates pavement cell interdigitation in Arabidopsis thaliana

In multicellular plant organs, cell shape formation depends on molecular switches to transduce developmental or environmental signals and to coordinate cell‐to‐cell communication. Plants have a specific subfamily of the Rho GT Pase family, usually called Rho of Plants(ROP), which serve as a critical signal transducer involved in many cellular processes. In the last decade, important advances in the ROP‐mediated regulation of plant cell morphogenesis have been made by using Arabidopsis thaliana leaf and cotyledon pavement cells.Especially, the auxin‐ROP signaling networks have been demonstrated to control interdigitated growth of pavement cells to form jigsaw‐puzzle shapes. Here, we review findings related to the discovery of this novel auxin‐signaling mechanism at the cell surface. This signaling pathway is to a large extent independent of the well‐known Transport Inhibitor Response(TIR)–Auxin Signaling F‐Box(AFB) pathway, and instead requires Auxin Binding Protein 1(ABP1) interaction with the plasma membrane‐localized, transmembrane kinase(TMK) receptor‐like kinase to regulate ROP proteins. Once activated, ROP influences cytoskeletal organization and inhibits endocytosis of the auxin transporter PIN1. The present review focuses on ROP signaling and its self‐organizing feature allowing ROP proteins to serve as a bustling signal decoder and integrator for plant cell morphogenesis.     

Phosphorylation-dependent differential regulation of plant growth, cell death, and innate immunity by the regulatory receptor-like kinase BAK1

Plants rely heavily on receptor-like kinases (RLKs) for perception and integration of external and internal stimuli. The Arabidopsis regulatory leucine-rich repeat RLK (LRR-RLK) BAK1 is involved in steroid hormone responses, innate immunity, and cell death control. Here, we describe the differential regulation of three different BAK1-dependent signaling pathways by a novel allele of BAK1, bak1-5. Innate immune signaling mediated by the BAK1-dependent RKs FLS2 and EFR is severely compromised in bak1-5 mutant plants. However, bak1-5 mutants are not impaired in BR signaling or cell death control. We also show that, in contrast to the RD kinase BRI1, the non-RD kinases FLS2 and EFR have very low kinase activity, and we show that neither was able to trans-phosphorylate BAK1 in vitro. Furthermore, kinase activity for all partners is completely dispensable for the ligand-induced heteromerization of FLS2 or EFR with BAK1 in planta, revealing another pathway specific mechanistic difference. The specific suppression of FLS2- and EFR-dependent signaling in bak1-5 is not due to a differential interaction of BAK1-5 with the respective ligand-binding RK but requires BAK1-5 kinase activity. Overall our results demonstrate a phosphorylation-dependent differential control of plant growth, innate immunity, and cell death by the regulatory RLK BAK1, which may reveal key differences in the molecular mechanisms underlying the regulation of ligand-binding RD and non-RD RKs.     

Characterization of a pollen-expressed receptor-like kinase gene of Petunia inflata and the activity of its encoded kinase.

From a pollen tube cDNA library of Petunia inflata, we isolated clones encoding a protein with structural features and biochemical properties characteristic of receptor-like kinases. It was designated PRK1 for pollen receptor-like kinase 1. The cytoplasmic domain of PRK1 is highly similar to the kinase domains of other plant receptor-like kinases and contains nearly all of the conserved amino acids for serine/threonine kinases. The extracellular domain of PRK1 contains leucine-rich repeats as found in some other plant receptor-like kinases, but overall its sequence in this region does not share significant similarity. Characterization of a gene encoding PRK1 revealed the presence of two introns. During pollen development, PRK1 mRNA was first detected in anthers containing mostly binucleate microspores; it reached the highest level of mature pollen and remained at a high level in in vitro-germinated pollen tubes. The recombinant cytoplasmic domain of PRK1 autophosphorylated on serine and tyrosine, suggesting that PRK1 may be a dual-specificity kinase. Monospecific immune serum to the recombinant extracellular domain of PRK1 detected a 69-kD protein in microsomal membranes of pollen and pollen tubes. The characteristics of PRK1 suggest that it may play a role in signal transduction events during pollen development and/or pollination.     

非豆科植物共生受体样蛋白激酶研究进展

植物根部能够与微生物形成相互依存、互惠互利的共生关系,非豆科植物根系主要与内生真菌形成菌根的共生体。共生受体样蛋白激酶(symbiosis receptor-like kinase,SYMRK)是植物识别菌根真菌诱导而产生的特异分子,它的蛋白结构由三个部分组成,即包含3个富含亮氨酸重复序列(LRRs)的胞外受体结合域、跨膜区和胞内蛋白激酶域。Symrk是控制共生形成的一个关键组分,该基因所编码的蛋白在植物识别和应答菌根真菌早期信号转导途径中是必需的。对Symrk基因的研究为进一步弄清植物-真菌共生的功能和作用机理打下了坚实的基础。     

生长素调控植物气孔发育的研究进展

气孔是分布于植物表皮由保卫细胞围成的小孔, 是植物体与外界环境进行水分和气体交换的重要通道, 通过影响光合作用、蒸腾作用及一系列生物学过程来促进植物适应环境的变化。生长素是最早被发现的植物激素, 在植物生长发育中发挥重要作用。近年来的研究表明, 生长素通过载体蛋白-TIR1/AFB受体-AUXIN/IAA-ARFs信号通路, 调控STOMAGEN的表达; 之后, 经STOMAGEN-类LRR受体蛋白激酶ERf-MAPKs级联反应激酶-SPCH转录因子信号通路, 启动气孔的发育进程。EPF1、EPF2和类LRR受体蛋白激酶TMM不是该过程的必需元件。生长素对气孔的调控受光信号影响, 光信号通路组分E3泛素连接酶COP1位于MAPKs激酶的上游, 参与气孔的发育调控。     

Diverse roles of SERK family genes in plant growth,development and defense response

Plant receptor-like protein kinases (RLKs) are transmembrane proteins with an extracellular domain and an intracellular kinase domain, which enable plant perceiving diverse extracellular stimuli to trigger the intracellular signal transduction. The somatic embryogenesis receptor kinases (SERKs) code the leucine-rich-repeat receptor-like kinase (LRR-RLK), and have been demonstrated to associate with multiple ligand-binding receptors to regulate plant growth, root development, male fertility, stomatal development and movement, and immune responses. Here, we focus on the progress made in recent years in understanding the versatile functions of Arabidopsis SERK proteins, and review SERK proteins as co-receptor to perceive different endogenous and environmental cues in different signaling pathway, and discuss how the kinase activity of SERKs is regulated by various modification.     

Identification, Expression and Functional Analysis of a Receptor-like Cytoplasmic Kinase, OsRLCK1, in Rice

Pollination involves a series of complex cellular interactions and signal transduction events. Numerous reports have suggested a central role for protein kinases in pollen germination and pollen tube growth and a large number of receptor-like kinases have been detected exclusively in pollen in higher plants. However, few are well characterized, especially for the receptor-like cytoplasmic kinases. Here we report a receptor-like kinase gene, OsRLCK1, which belongs to the receptor-like cytoplasmic kinase Ⅷ subfamily. Real-time quantitative polymerase chain reaction analysis and whole mount RNA in situ hybridization showed that OsRLCK1 is a pollen-specific gene and expressed only in the mature pollen. When expressed in the onion epidermal cells, the OsRLCK1-GFP fusion protein was diffused throughout the cell, indicating its cytoplasmic and nuclear localization. The Maltose Binding Protein-OsRLCK1 recombinant protein was found to be capable of autophosphorylation on threonine residue, showing that it encodes a functional kinase. These results suggest that OsRLCK1 is likely to play a role in a signaling pathway associated with pollen performance during pollination in rice.     

Brassinosteroid Signal Transduction: A Mix of Conservation and Novelty

Brassinosteroids (BRs) are a unique class of plant steroids that are structurally similar to animal steroid hormones and play important roles in plant growth and development. Unlike the animal steroids, which bind to classical intracellular steroid receptors that directly modulate gene activities after translocation into the nucleus, the plant steroids rely on transmembrane receptor kinases to activate a phosphorylation cascade to regulate gene expression. Recent genetic and biochemical studies have identified several critical BR signaling components and revealed a striking mechanistic similarity between the plant steroid signaling pathway and several well-studied animal signaling cascades involving a receptor kinase and glycogen synthase kinase 3 (GSK3). A working model for BR signal transduction proposes that BR initiates its signaling pathway by promoting heterodimerization of two transmembrane receptor-like kinases at the cell surface, leading to inhibition of a GSK3 kinase and subsequent stabilization and nuclear accumulation of two GSK3 substrates that regulate BR-responsive genes. Such a simple model provides a framework for continued investigation of molecular mechanism(s) of plant steroid signaling.     

Rice A20/AN1 zinc-finger containing stress-associated proteins (SAP1/11) and a receptor-like cytoplasmic kinase (OsRLCK253) interact via A20 zinc-finger and confer abiotic stress tolerance in transgenic Arabidopsis plants

? The inbuilt mechanisms of plant survival have been exploited for improving tolerance to abiotic stresses. Stress-associated proteins (SAPs), containing A20/AN1 zinc-finger domains, confer abiotic stress tolerance in different plants, however, their interacting partners and downstream targets remain to be identified. ? In this study, we have investigated the subcellular interactions of rice SAPs and their interacting partner using yeast two-hybrid and fluorescence resonance energy transfer (FRET) approaches. Their efficacy in improving abiotic stress tolerance was analysed in transgenic Arabidopsis plants. Regulation of gene expression by genome-wide microarray in transgenics was used to identify downstream targets. ? It was found that the A20 domain mediates the interaction of OsSAP1 with self, its close homolog OsSAP11 and a rice receptor-like cytoplasmic kinase, OsRLCK253. Such interactions between OsSAP1/11 and with OsRLCK253 occur at nuclear membrane, plasma membrane and in nucleus. Functionally, both OsSAP11 and OsRLCK253 could improve the water-deficit and salt stress tolerance in transgenic Arabidopsis plants via a signaling pathway affecting the expression of several common endogenous genes. ? Components of a novel stress-responsive pathway have been identified. Their stress-inducible expression provided the protection against yield loss in transgenic plants, indicating the agronomic relevance of OsSAP11 and OsRLCK253 in conferring abiotic stress tolerance.     

SbRLK1, a receptor-like protein kinase of Sorghum bicolor (L.) Moench that is expressed in mesophyll cells

To study the metabolic interactions between mesophyll and bundle-sheath cells of C₄ plants, protein kinases possibly involved in the regulatory processes and signal transduction pathways have been cloned and characterized. A receptor-like protein kinase (RLK) cDNA clone from the C₄ plant Sorghum bicolor (L.) Moench has been identified. The deduced protein was designated SbRLK1 for receptor-like protein kinase from S. bicolor. The putative cytoplasmic domain of SbRLK1 contains all amino acids that are characteristic of protein kinases. The extracellular domain contains five leucine-rich repeats. The mRNA of the SbRLK1 gene accumulated to much higher levels in mesophyll cells than in the bundle-sheath and was almost undetectable in roots. This expression pattern indicates that SbRLK1 might be involved in the regulation of specific processes in mesophyll cells. Received: 13 August 1998 / Accepted: 22 December 1998     

Water-deficit-responsive proteins in maritime pine

We have isolated three receptor-like kinase cDNAs from an Arabidopsis flower cDNA library by PCR using degenerate oligonucleotide primers for conserved domains of protein kinases. Cloning and sequencing of the full-length cDNAs, designated RKF1 to 3 (receptor-like kinase in flowers), showed that the putative extracellular domain of the RKF1 protein contains 13 tandem repeats of leucine-rich sequences and those of RKF2 and RKF3 have no significant homology with other plant sequences. RNA blot analysis revealed that the RKF1 mRNA is highly expressed in stamens while RKF2 and RKF3 mRNAs are present at low levels in all organs examined. In situ localization experiments indicated that the RKF1 mRNA is detectable in early flower primordia and during stamen development. In addition, when fused to a GUS reporter gene, the RKF1 promoter directed high GUS expression in pollen grains. Recombinant RKF1, produced in Escherichia coli, was found to have kinase activity with serine/threonine specificity in vitro.