In two cases, mutations in the same brassinosteroid-related genes caused different phenotypes in japonica varieties Nipponbare and Taichung 65. The mutant phenotypes were less severe in the Taichung 65 background than...In two cases, mutations in the same brassinosteroid-related genes caused different phenotypes in japonica varieties Nipponbare and Taichung 65. The mutant phenotypes were less severe in the Taichung 65 background than in the Nipponbare background. Three newly isolated brassinosteroid-insensitive mutants (d61-1N, d61-11, and d61-12) derived from a Nipponbare mutant library were found to be alleles of d61, which represent defects in the OsBRI1 gene. Although the Nipponbare-derived mutant d61-1N had the same nucleotide substitution as the previously characterized Taichung 65-derived mutant d61-1T, these two mutants showed different phenotypes for plant stature, internode elongation pattern, and seed shape;in each case, d61-1N (in the Nipponbare genetic background) had the more severe mutant phenotype. Similar trends were seen for phenotypes caused by mutants of d2, a brassinosteroid biosynthesis gene. Consistent with these phenotypes, the expression of brassinosteroid-responsive genes was lower in the Nipponbare-derived mutants. These results can be explained by our findings that feed-forward up-regulation of OsBRI1 did not occur in the Nipponbare-derived mutants and that an mPing transposon is inserted into the promoter region of Nipponbare OsBRI1. Based on these results, we conclude that the expression of OsBRI1, especially its feed-forward up-regulation, is misregulated in wild-type Nipponbare and in brassinosteroid-related mutants in a Nipponbare genetic background. Although Nipponbare is a model rice genotype, it can be categorized as an OsBRI1 mutant that has reduced sensitivity to brassinosteroid.展开更多
Brassinosteroid （BR） binding activates the receptor kinase BRI1 by inducing heterodimerization with its co- receptor kinase BAK1; however, the mechanisms that reversibly inactivate BRI1 remain unclear. Here we show ...Brassinosteroid （BR） binding activates the receptor kinase BRI1 by inducing heterodimerization with its co- receptor kinase BAK1; however, the mechanisms that reversibly inactivate BRI1 remain unclear. Here we show that cytoplasm-localized protein phosphatase 2A （PP2A） B＇ regulatory subunits interact with BRI1 to mediate its dephosphorylation and inactivation. Loss-of-function and overexpression experiments showed that a group of PP2A B＇ regulatory subunits, represented by B＇η, negatively regulate BR signaling by decreasing BRI1 phosphorylation. BR increases the expression levels of these B＇ subunits, and B/TI interacts preferentially with phosphorylated BRI1, suggesting that the dynamics of BR signaling are modu- lated by the PP2A-mediated feedback inactivation of BRI1. Compared with PP2A B＇α and B＇β, which promote BR responses by dephosphorylating the downstream transcription factor BZR1, the BRI1- inactivating B＇ subunits showed similar binding to BRI1 and BZR1 but distinct subcellular localization. Alteration of the nuclear/cytoplasmic localization of the B＇ subunits revealed that cytoplasmic PP2A de- phosphorylates BRI1 and inhibits the BR response, whereas nuclear PP2A dephosphorylates BZR1 and ac- tivates the BR response. Our findings not only identify the PP2A regulatory B subunits that mediate the binding and dephosphorylation of BRI1, but also demonstrate that the subcellular localization of PP2A specifies its substrate selection and distinct effects on BR signaling.展开更多
Leucine-rich repeat receptor-like kinases （LRR-RLKs） belong to a large group of cell surface proteins involved in many aspects of plant development and environmental responses in both monocots and dicots. Brassinost...Leucine-rich repeat receptor-like kinases （LRR-RLKs） belong to a large group of cell surface proteins involved in many aspects of plant development and environmental responses in both monocots and dicots. Brassinosteroid insensitive 1 （BRI1）, a member of the LRR X subfamily, was first identified through several forward genetic screenings for mutants insensitive to brassinosteroids （BRs）, which are a class of plant-specific steroid hormones. Since its identification, BRI1 and its homologs had been proved as receptors perceiving BRs and initiating BR signaling. The co-receptor BRIl-associated kinase 1 and its homologs, and other BRI1 interacting proteins such as its inhibitor BRI1 kinase inhibitor I （BKI1） were identified by genetic andbiochemical approaches. The detailed mechanisms of BR perception by BRI1 and the activation of BRI1 receptor complex have also been elucidated. Moreover, several mechanisms for termination of the activated BRI1 signaling were also discovered. In this review, we will focus on the recent advances on the mechanism of BRI1 phosphorylation and activation, the regulation of its receptor complex, the structure basis of BRI1 ectodomain and BR recognition, its direct substrates, and the termination of the activated BRI1 receptor complex.展开更多
The endoplasmic reticulum-associated degradation （ERAD） is a highly conserved mechanism to remove mis- folded membrane/secretory proteins from the endoplasmic reticulum （ER）. While many of the individual component...The endoplasmic reticulum-associated degradation （ERAD） is a highly conserved mechanism to remove mis- folded membrane/secretory proteins from the endoplasmic reticulum （ER）. While many of the individual components of the ERAD machinery are well characterized in yeast and mammals, our knowledge of a plant ERAD process is rather limited. Here, we report a functional study of an Arabidopsis homolog （AtOS9） of an ER luminal lectin Yos9 （OS-9 in mammals） that recognizes a unique asparagine-linked glycan on misfolded proteins. We discovered that AtOS9 is an ER-Iocalized glyco- protein that is co-expressed with many known/predicted ER chaperones. AT-DNA insertional atos9-t mutation blocks the degradation of a structurally imperfect yet biochemically competent brassinosteroid （BR） receptor bril-9, causing its increased accumulation in the ER and its consequent leakage to the cell surface responsible for restoring the BR sensitivity and suppressing the dwarfism of the bril-9 mutant. In addition, we identified a missense mutation in AtOS9 in a recently discovered ERAD mutant ems-rnutagenized bril suppressor 6 （ebs6-1）. Moreover, we showed that atos9-t also inhibits the ERAD of bril-5, another ER-retained BR receptor, and a misfolded EFR, a BRIl-like receptor for the bacterial translation elongation factor EF-Tu. Furthermore, we found that AtOS9 interacted biochemically and genetically with EBS5, an Arabidopsis homolog of the yeast Hrd3/mammalian SellL known to collaborate with Yos9/OS-9 to select ERAD clients. Taken together, our results demonstrated a functional role of AtOS9 in a plant ERAD process that degrades misfolded receptor-like kinases.展开更多
Plants utilize plasma membrane-localized receptor-like kinases （RLKs） to sense extracellular signals to coordinate growth, development, and innate immune responses. BAK1 regulates multiple signaling pathways acting ...Plants utilize plasma membrane-localized receptor-like kinases （RLKs） to sense extracellular signals to coordinate growth, development, and innate immune responses. BAK1 regulates multiple signaling pathways acting as a co-receptor of several distinct ligand-binding RLKs. It has been debated whether BAK1 serves as an essential regulatory component or only a signal amplifier without pathway specificity. This issue has been clarified recently. Genetic and structural analyses indicated that BAK1 and its homologs play indispensible roles in mediating brassinosteroid （BR） signaling pathway by directly perceiving the ligand BR and activating the receptor of BR, BRII. The mechanism revealed by these studies now serves as a paradigm for how a pair of RLKs can function together in ligand binding and subsequent initiation of signaling.展开更多
文摘In two cases, mutations in the same brassinosteroid-related genes caused different phenotypes in japonica varieties Nipponbare and Taichung 65. The mutant phenotypes were less severe in the Taichung 65 background than in the Nipponbare background. Three newly isolated brassinosteroid-insensitive mutants (d61-1N, d61-11, and d61-12) derived from a Nipponbare mutant library were found to be alleles of d61, which represent defects in the OsBRI1 gene. Although the Nipponbare-derived mutant d61-1N had the same nucleotide substitution as the previously characterized Taichung 65-derived mutant d61-1T, these two mutants showed different phenotypes for plant stature, internode elongation pattern, and seed shape;in each case, d61-1N (in the Nipponbare genetic background) had the more severe mutant phenotype. Similar trends were seen for phenotypes caused by mutants of d2, a brassinosteroid biosynthesis gene. Consistent with these phenotypes, the expression of brassinosteroid-responsive genes was lower in the Nipponbare-derived mutants. These results can be explained by our findings that feed-forward up-regulation of OsBRI1 did not occur in the Nipponbare-derived mutants and that an mPing transposon is inserted into the promoter region of Nipponbare OsBRI1. Based on these results, we conclude that the expression of OsBRI1, especially its feed-forward up-regulation, is misregulated in wild-type Nipponbare and in brassinosteroid-related mutants in a Nipponbare genetic background. Although Nipponbare is a model rice genotype, it can be categorized as an OsBRI1 mutant that has reduced sensitivity to brassinosteroid.
文摘Brassinosteroid （BR） binding activates the receptor kinase BRI1 by inducing heterodimerization with its co- receptor kinase BAK1; however, the mechanisms that reversibly inactivate BRI1 remain unclear. Here we show that cytoplasm-localized protein phosphatase 2A （PP2A） B＇ regulatory subunits interact with BRI1 to mediate its dephosphorylation and inactivation. Loss-of-function and overexpression experiments showed that a group of PP2A B＇ regulatory subunits, represented by B＇η, negatively regulate BR signaling by decreasing BRI1 phosphorylation. BR increases the expression levels of these B＇ subunits, and B/TI interacts preferentially with phosphorylated BRI1, suggesting that the dynamics of BR signaling are modu- lated by the PP2A-mediated feedback inactivation of BRI1. Compared with PP2A B＇α and B＇β, which promote BR responses by dephosphorylating the downstream transcription factor BZR1, the BRI1- inactivating B＇ subunits showed similar binding to BRI1 and BZR1 but distinct subcellular localization. Alteration of the nuclear/cytoplasmic localization of the B＇ subunits revealed that cytoplasmic PP2A de- phosphorylates BRI1 and inhibits the BR response, whereas nuclear PP2A dephosphorylates BZR1 and ac- tivates the BR response. Our findings not only identify the PP2A regulatory B subunits that mediate the binding and dephosphorylation of BRI1, but also demonstrate that the subcellular localization of PP2A specifies its substrate selection and distinct effects on BR signaling.
基金supported by the National Natural Science Foundation of China to X.W.(91117005 and 30925020)National Basic Research Program of China to X.W.(2012CB114300)+1 种基金Key Project of Shanghai Science and Technology Committee to X.W.(10JC1400800)Program of Shanghai Subject Chief Scientist to X.W.(11XD1400700)
文摘Leucine-rich repeat receptor-like kinases （LRR-RLKs） belong to a large group of cell surface proteins involved in many aspects of plant development and environmental responses in both monocots and dicots. Brassinosteroid insensitive 1 （BRI1）, a member of the LRR X subfamily, was first identified through several forward genetic screenings for mutants insensitive to brassinosteroids （BRs）, which are a class of plant-specific steroid hormones. Since its identification, BRI1 and its homologs had been proved as receptors perceiving BRs and initiating BR signaling. The co-receptor BRIl-associated kinase 1 and its homologs, and other BRI1 interacting proteins such as its inhibitor BRI1 kinase inhibitor I （BKI1） were identified by genetic andbiochemical approaches. The detailed mechanisms of BR perception by BRI1 and the activation of BRI1 receptor complex have also been elucidated. Moreover, several mechanisms for termination of the activated BRI1 signaling were also discovered. In this review, we will focus on the recent advances on the mechanism of BRI1 phosphorylation and activation, the regulation of its receptor complex, the structure basis of BRI1 ectodomain and BR recognition, its direct substrates, and the termination of the activated BRI1 receptor complex.
基金This work was partly supported by grants from National Institutes of Health （GM060519） and National Science Foundation （IOS 1121496） to J.L.
文摘The endoplasmic reticulum-associated degradation （ERAD） is a highly conserved mechanism to remove mis- folded membrane/secretory proteins from the endoplasmic reticulum （ER）. While many of the individual components of the ERAD machinery are well characterized in yeast and mammals, our knowledge of a plant ERAD process is rather limited. Here, we report a functional study of an Arabidopsis homolog （AtOS9） of an ER luminal lectin Yos9 （OS-9 in mammals） that recognizes a unique asparagine-linked glycan on misfolded proteins. We discovered that AtOS9 is an ER-Iocalized glyco- protein that is co-expressed with many known/predicted ER chaperones. AT-DNA insertional atos9-t mutation blocks the degradation of a structurally imperfect yet biochemically competent brassinosteroid （BR） receptor bril-9, causing its increased accumulation in the ER and its consequent leakage to the cell surface responsible for restoring the BR sensitivity and suppressing the dwarfism of the bril-9 mutant. In addition, we identified a missense mutation in AtOS9 in a recently discovered ERAD mutant ems-rnutagenized bril suppressor 6 （ebs6-1）. Moreover, we showed that atos9-t also inhibits the ERAD of bril-5, another ER-retained BR receptor, and a misfolded EFR, a BRIl-like receptor for the bacterial translation elongation factor EF-Tu. Furthermore, we found that AtOS9 interacted biochemically and genetically with EBS5, an Arabidopsis homolog of the yeast Hrd3/mammalian SellL known to collaborate with Yos9/OS-9 to select ERAD clients. Taken together, our results demonstrated a functional role of AtOS9 in a plant ERAD process that degrades misfolded receptor-like kinases.
基金supported by the grants from the National Natural Science Foundation of China to J.L.(91117008 and 90917019)National Basic Research Program of China to J.L.(2011CB915401)Fundamental Research Funds for the Central Universities to S.X.(lzujbky-2009-35)
文摘Plants utilize plasma membrane-localized receptor-like kinases （RLKs） to sense extracellular signals to coordinate growth, development, and innate immune responses. BAK1 regulates multiple signaling pathways acting as a co-receptor of several distinct ligand-binding RLKs. It has been debated whether BAK1 serves as an essential regulatory component or only a signal amplifier without pathway specificity. This issue has been clarified recently. Genetic and structural analyses indicated that BAK1 and its homologs play indispensible roles in mediating brassinosteroid （BR） signaling pathway by directly perceiving the ligand BR and activating the receptor of BR, BRII. The mechanism revealed by these studies now serves as a paradigm for how a pair of RLKs can function together in ligand binding and subsequent initiation of signaling.
