泛素化修饰调控脱落酸介导的信号途径

泛素化修饰是一种重要的蛋白质翻译后修饰,通过调节蛋白的活性和稳定性等影响其功能的发挥,在真核生物的生命过程中具有非常重要的作用。泛素化修饰通过精细地调控植物激素脱落酸(abscisic acid, ABA)的合成和信号转导过程的关键因子,影响植物对ABA的响应,参与植物生长发育过程及对干旱、盐和冷胁迫等不良环境的应答。本文概述了植物中泛素化修饰的相关组分(包括泛素连接酶E3、泛素结合酶E2、26S蛋白酶体)和内膜运输相关蛋白,以及这些蛋白调控ABA合成和信号转导过程的最新研究进展,提出该研究领域需要解决的新问题,以期为相关领域的科研人员进一步了解翻译后修饰如何调控激素信号的转导途径提供参考。     

An efficient system to detect protein ubiquitination by agroinfiltration in Nicotiana benthamiana

The ubiquitination proteasome pathway has been demonstrated to regulate all plant developmental and signaling processes. E3 ligase/substrate‐specific interactions and ubiquitination play important roles in this pathway. However, due to technical limitations only a few instances of E3 ligase–substrate binding and protein ubiquitination in plants have been directly evidenced. An efficient in vivo and in vitro ubiquitination assay was developed for analysis of protein ubiquitination reactions by agroinfiltration expression of both substrates and E3 ligases in Nicotiana benthamiana. Using a detailed analysis of the well‐known E3 ligase COP1 and its substrate HY5, we demonstrated that this assay allows for fast and reliable detection of the specific interaction between the substrate and the E3 ligase, as well as the effects of MG132 and substrate ubiquitination and degradation. We were able to differentiate between the original and ubiquitinated forms of the substrate in vivo with antibodies to ubiquitin or to the target protein. We also demonstrated that the substrate and E3 ligase proteins expressed by agroinfiltration can be applied to analyze ubiquitination in in vivo or in vitro reactions. In addition, we optimized the conditions for different types of substrate and E3 ligase expression by supplementation with the gene‐silencing suppressor p19 and by time‐courses of sample collection. Finally, by testing different protein extraction buffers, we found that different types of buffer should be used for different ubiquitination analyses. This method should be adaptable to other protein modification studies.     

Regulation of abiotic stress signal transduction by E3 ubiquitin ligases in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Ubiquitination is a unique protein degradation system utilized by eukaryotes to efficiently degrade detrimental cellular proteins and control the entire pool of regulatory components. In plants, adaptation in response to various abiotic stresses can be achieved through ubiquitination and the resulting degradation of components specific to these stress signalings. Arabidopsis has more than 1,400 E3 enzymes, indicating E3 ligase acts as a main determinant of substrate specificity. However, as only a minority of E3 ligases related to abiotic stress signaling have been studied in Arabidopsis, the further elucidation of the biological roles and related substrates of newly identified E3 ligases is essential in order to clarify the functional relationship between abiotic stress and E3 ligases. Here, we review the current knowledge and future prospects of the regulatory mechanism and role of several E3 ligases involved in abiotic stress signal transduction in Arabidopsis. As another potential approach to understand how ubiquitination is involved in such signaling, we also briefly introduce factors that regulate the activity of cullin in multisubunit E3 ligase complexes.     

The E3 ubiquitin ligase WVIP2 highlights the versatility of protein ubiquitination

Plant cells regulate many cellular processes controlling the half-life of critical proteins through ubiquitination. Previously, we characterized two interacting RING-type E3 ubiquitin ligases of Triticum durum, TdRF1 and WVIP2. We revealed their role in tolerance to dehydration, and existing knowledge about their partners also indicated their involvement in the regulation of some aspects of plant development. Here we located WVIP2 in the regulation of the ABA signaling, based on sequence similarities. Further we acquired general evidence about the versatility of ubiquitination in plant cells. A protein can be target of different E3 ligases for a perfect tuning of its abundance as well as the same E3 ligase can ubiquitinate different and unrelated proteins, thus representing a cross-connections between different signaling pathways for a global coordination of cellular processes.     

The regulation of TGF-β/SMAD signaling by protein deubiquitination

Transforming growth factor-β (TGF-β) members are key cytokines that control embryogenesis and tissue homeostasis via transmembrane TGF-β type II (TβR II) and type I (TβRI) and serine/threonine kinases receptors. Aberrant activation of TGF-β signaling leads to diseases, including cancer. In advanced cancer, the TGF-β/SMAD pathway can act as an oncogenic factor driving tumor cell invasion and metastasis, and thus is considered to be a therapeutic target. The activity of TGF-β/SMAD pathway is known to be regulated by ubiquitination at multiple levels. As ubiquitination is reversible, emerging studies have uncovered key roles for ubiquitin-removals on TGF-β signaling components by deubiquitinating enzymes (DUBs). In this paper, we summarize the latest findings on the DUBs that control the activity of the TGF-β signaling pathway. The regulatory roles of these DUBs as a driving force for cancer progression as well as their underlying working mechanisms are also discussed.     

E3 ubiquitin ligases and abscisic acid signaling

The ubiquitin proteasome system is involved in the regulation of nearly every aspect of plant growth and development. Protein ubiquitination involves the covalent attachment of ubiquitin to target proteins through a cascade catalyzed by three enzymes known as E1, E2 and E3. E3s are of particular interest as they confer substrate specificity during ubiquitination through their diverse substrate recognition domains. Recently, a number of E3s have been identified that actively participate in abscisic acid hormone biology, including regulation of biosynthesis, de-repression or activation of abscisic acid response and degradation of signaling components. In this review, we summarize recent exciting studies of the different types of E3s that target specific mediators of abscisic acid signaling or affect the plants response to the hormone.Key words: abscisic acid, E3 ubiquitin ligase, proteasome, ubiquitinationPost-translational control of protein degradation by the ubiquitin proteasome system (UPS) is a highly regulated process essential for the proper growth and development of all eukaryotes through removing abnormal proteins and most short-lived regulatory proteins.^1,2 Plants utilize the UPS to alter their proteome to mediate cellular changes required for growth, development and responses to biotic and abiotic stress. Plants also rely a great deal on hormones to induce changes in growth and development in response to a wide range of environmental stimuli. Hormone biosynthesis, perception, signaling and response can be exquisitely regulated through modulating protein levels via the UPS. Regulation of the abscisic acid (ABA) signaling pathway, like auxin, gibberellin, jasmonate and ethylene, have been linked to UPS components with the application of biochemical, genetic and genomic approaches.^3–5 Although some aspects of ABA signaling have been elucidated, the involvement of the UPS, especially E3 ubiquitin ligases, help us gain further insight into the entire network of ABA signal transduction. In this review we focus on recently identified E3s that play a variety of roles in ABA signaling. A number of articles are available that provide a comprehensive review of the role of E3 ligases in the biosynthesis, perception and signaling by other hormones such as auxin and ethylene.^3–5     

ABA信号通路的起源与进化

植物激素脱落酸（abscisic acid,ABA）在植物的生长、发育和胁迫反应方面起重要的调控作用,其信号转导通路由4个核心组分共同组成一个双重负调控系统（PYR/PYL/RCAR—| PP2C—| SnRK2—ABF/AREB）,调控ABA应答反应。本文在综述和分析ABA信号通路4个核心组分的起源与进化的基础上,初步提出ABA信号通路的起源与进化路径：A类PP2C、第Ⅲ亚类SnRK2以及转录因子AREB/ABF在水生植物轮藻中已经进化产生,当陆生植物进化产生ABA受体PYR/PYL/RCAR后,即与其它3个组分形成完整的ABA信号通路。在植物进化过程中,ABA信号通路4个核心组分各家族成员的数量（亚类）呈递增趋势。     

MAPK级联途径参与ABA信号转导调节的植物生长发育过程

植物激素ABA参与调控植物生长发育和生理代谢以及多种胁迫应答过程,促分裂原活化蛋白激酶（MAPK）级联途径应答于多种生物和非生物胁迫,广泛参与调控植物的生长发育。MAPK级联途径与ABA信号转导协同作用参与调控植物种子萌发、气孔运动和生长发育,本文主要归纳了植物中受ABA调控激活的MAPK级联途径成员,阐述了它们参与ABA信号转导调控植物生理反应和生长发育的过程,并对MAPK级联途径与ABA信号转导的研究方向作出了展望,指出对MAPK下游底物的筛选是完善MAPK级联途径的重要组成部分。     

蛋白质翻译后修饰在ABA信号转导中的作用

脱落酸(ABA)是植物生长发育和逆境适应过程中非常关键的植物激素。植物响应ABA信号转导过程由信号识别、转导及响应级联完成, 其中心转导途径由ABA受体RCAR/PYR/PYLs、磷酸酶PP2Cs、激酶SnRK2s、转录因子和离子通道蛋白构成。蛋白磷酸化、泛素化、类泛素化和氧化还原等翻译后修饰在ABA转导途径中起重要作用。该文综述了翻译后修饰在ABA信号转导中的作用。     

蛋白质翻译后修饰在ABA信号转导中的作用

脱落酸(ABA)是植物生长发育和逆境适应过程中非常关键的植物激素。植物响应ABA信号转导过程由信号识别、转导及响应级联完成, 其中心转导途径由ABA受体RCAR/PYR/PYLs、磷酸酶PP2Cs、激酶SnRK2s、转录因子和离子通道蛋白构成。蛋白磷酸化、泛素化、类泛素化和氧化还原等翻译后修饰在ABA转导途径中起重要作用。该文综述了翻译后修饰在ABA信号转导中的作用。     

Regulation of Hedgehog signaling by ubiquitination

The Hedgehog (Hh) signaling pathway plays crucial roles both in embryonic development and in adult stem cell function. The timing, duration and location of Hh signaling activity need to be tightly controlled. Abnormalities of Hh signal transduction lead to birth defects or malignant tumors. Recent data point to ubiquitination-related posttranslational modifications of several key Hh pathway components as an important mechanism of regulation of the Hh pathway. Here we review how ubiquitination regulates the localization, stability and activity of the key Hh signaling components.     

ABA inhibits myristoylation and induces shuttling of the RGLG1 E3 ligase to promote nuclear degradation of PP2CA

Hormone‐ and stress‐induced shuttling of signaling or regulatory proteins is an important cellular mechanism to modulate hormone signaling and cope with abiotic stress. Hormone‐induced ubiquitination plays a crucial role to determine the half‐life of key negative regulators of hormone signaling. For ABA signaling, the degradation of clade‐A PP 2Cs, such as PP 2 CA or ABI 1, is a complementary mechanism to PYR / PYL / RCAR ‐mediated inhibition of PP 2C activity. ABA promotes the degradation of PP 2 CA through the RGLG 1 E3 ligase, although it is not known how ABA enhances the interaction of RGLG 1 with PP 2 CA given that they are predominantly found in the plasma membrane and the nucleus, respectively. We demonstrate that ABA modifies the subcellular localization of RGLG 1 and promotes nuclear interaction with PP 2 CA . We found RGLG 1 is myristoylated in vivo , which facilitates its attachment to the plasma membrane. ABA inhibits the myristoylation of RGLG 1 through the downregulation of N‐myristoyltransferase 1 ( NMT 1 ) and promotes nuclear translocation of RGLG 1 in a cycloheximide‐insensitive manner. Enhanced nuclear recruitment of the E3 ligase was also promoted by increasing PP 2 CA protein levels and the formation of RGLG 1–receptor–phosphatase complexes. We show that RGLG 1 ^Gly2Ala mutated at the N‐terminal myristoylation site shows constitutive nuclear localization and causes an enhanced response to ABA and salt or osmotic stress. RGLG 1/5 can interact with certain monomeric ABA receptors, which facilitates the formation of nuclear complexes such as RGLG 1– PP 2 CA – PYL 8. In summary, we provide evidence that an E3 ligase can dynamically relocalize in response to both ABA and increased levels of its target, which reveals a mechanism to explain how ABA enhances RGLG 1– PP 2 CA interaction and hence PP 2 CA degradation.     

The role of ubiquitination and deubiquitination in tumor invasion and metastasis

Ubiquitination is vital for multiple cellular processes via dynamic modulation of proteins related to cell growth, proliferation, and survival. Of the ubiquitination system components, E3 ubiquitin ligases and deubiquitinases have the most prominent roles in modulating tumor metastasis. This review will briefly summarize the observations and underlying mechanisms of multiple E3 ubiquitin ligases and deubiquitinases to regulate tumor metastasis. Further, we will discuss the relationship and importance between ubiquitination components and tumor progression.     

Ubiquitination in NB-LRR-mediated immunity

As a common protein modification, ubiquitination is used for regulating the fate of protein targets, notably in terms of stability. In recent years, it has emerged to play key roles in the regulation of plant defense responses. Given its flexibility and critical roles in signaling, primarily in the control of protein turnover, ubiquitination is probably targeting many major immune regulators for modification or degradation. In this review, we summarize the latest findings on how different components of the ubiquitination pathway are involved in NB-LRR R protein-mediated immunity.     

The PP2C-SnRK2 complex: The central regulator of an abscisic acid signaling pathway

The phytohormone abscisic acid (ABA), an important bioactive compound in plants, is implicated in several essential processes such as development and the abiotic stress response. Many components have been reported to have roles in these processes. Although 2C-type protein phosphatases (PP2C) and SNF1-related protein kinases2 (SnRK2) family are known to be important signal mediators, the molecular mechanisms by which these components regulate the ABA signaling pathway have not been elucidated. Recent identification of soluble ABA receptors, PYR/PYL/RCAR, has provided a major breakthrough in understanding the signaling mechanisms of ABA and revealed the importance of PP2Cs. In addition, the physical, biochemical and physiological connections between PP2C and SnRK2 have been clearly demonstrated. Taken together, the molecular basis of the major ABA signaling pathway has been established, from perception to gene expression. In this addendum, we discuss this emerging ABA signaling pathway, which has a conventional protein phosphorylation/dephosphorylation regulatory circuit and consider its physiological and functional relevance.Key words: ABA receptor, abscisic acid, PP2C, signal transduction, SnRK2, plant hormone, phosphoarylation     

PYR/PYL/RCAR蛋白介导植物ABA的信号转导

脱落酸(ABA)在各个植物生长发育阶段以及植物对生物与非生物胁迫的响应过程中都发挥着重要的作用。最近研究表明,在ABA信号转导途径中有3种核心组份:ABA受体PYR/PYL/RCAR蛋白、负调控因子2C类蛋白磷酸酶(PP2C)和正调控因子SNF1相关的蛋白激酶2(SnRK2),它们共同组成了一个双重负调控系统——PYR/PYL/RCAR—|PP2C—|SnRK2来调控ABA信号转导及其下游反应,且3种核心组份在植物体内的结合方式受时空和生化等因素的影响,通过特定组合形成的ABA信号转导复合体介导特定的ABA信号反应。文章就PYR/PYL/RCAR蛋白介导的植物ABA信号识别与转导途径的分子基础及其调控机制,以及PYR/PYL/RCAR—PP2C—SnRK2参与的ABA信号调控网络等研究进展做一概述,并对该领域今后的研究进行了展望。