Mehali Mitra , Samrat Banerjee , Bhagath Kumar Palaka , Swarup Roy Choudhury , Sujit Roy
{"title":"拟南芥中的核 WD40 重复蛋白 PRL1 可调节 MYB4 转录因子的稳定性","authors":"Mehali Mitra , Samrat Banerjee , Bhagath Kumar Palaka , Swarup Roy Choudhury , Sujit Roy","doi":"10.1016/j.cpb.2024.100341","DOIUrl":null,"url":null,"abstract":"<div><p>MYB4, a member of the R2R3-type subfamily of MYB transcription factor plays a crucial role in regulating the accumulation of UV-B absorbing phenylpropanoids in plants. UV-B exposure for a longer duration down-regulates the expression of <em>MYB4</em> gene in <em>Arabidopsis</em>. MYB4 protein represses its own expression by binding to its own promoter. However, at present practically nothing is known about the post-translational regulation of MYB4 protein <em>in vivo</em>. Here, we provide evidence that in <em>Arabidopsis</em> MYB4 protein is phosphorylated <em>in vivo</em> and is targeted by the ubiquitin-26S proteasome-dependent pathway. Immunoprecipitation, immunoblotting, and phosphoprotein staining experiments have revealed that both the accumulation pattern and phosphorylation of MYB4 increase in the light condition during the 24 hours time span under long-day conditions. Yeast two-hybrid and bimolecular fluorescence complementation assays have shown that MYB4 directly interacts with a nuclear WD40 repeat protein, PRL1 <em>in vivo</em>. Cell-free protein degradation assay in the absence and presence of proteasome inhibitor indicates that MYB4 is degraded in a ubiquitin proteasome-dependent manner. Furthermore, analyses of MYB4 protein accumulation levels in transgenic <em>atmyb4–1</em> mutant line expressing <em>35</em> <em>S:AtMYB4</em> (<em>35</em> <em>S:AtMYB4-atmyb4–1</em>) and <em>atprl1–1</em> mutant line indicate that PRL1 regulate stability of MYB4 in <em>Arabidopsis</em>. Overall, our results provide important information on the possible mechanism of post-translational modification and regulation of stability of MYB4 protein in <em>Arabidopsis in vivo</em>.</p></div>","PeriodicalId":38090,"journal":{"name":"Current Plant Biology","volume":null,"pages":null},"PeriodicalIF":5.4000,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214662824000239/pdfft?md5=2e9b5c0a08002a2e3ecba581f77817ca&pid=1-s2.0-S2214662824000239-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A nuclear WD40 repeat protein PRL1 regulates stability of MYB4 transcription factor in Arabidopsis\",\"authors\":\"Mehali Mitra , Samrat Banerjee , Bhagath Kumar Palaka , Swarup Roy Choudhury , Sujit Roy\",\"doi\":\"10.1016/j.cpb.2024.100341\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>MYB4, a member of the R2R3-type subfamily of MYB transcription factor plays a crucial role in regulating the accumulation of UV-B absorbing phenylpropanoids in plants. UV-B exposure for a longer duration down-regulates the expression of <em>MYB4</em> gene in <em>Arabidopsis</em>. MYB4 protein represses its own expression by binding to its own promoter. However, at present practically nothing is known about the post-translational regulation of MYB4 protein <em>in vivo</em>. Here, we provide evidence that in <em>Arabidopsis</em> MYB4 protein is phosphorylated <em>in vivo</em> and is targeted by the ubiquitin-26S proteasome-dependent pathway. Immunoprecipitation, immunoblotting, and phosphoprotein staining experiments have revealed that both the accumulation pattern and phosphorylation of MYB4 increase in the light condition during the 24 hours time span under long-day conditions. Yeast two-hybrid and bimolecular fluorescence complementation assays have shown that MYB4 directly interacts with a nuclear WD40 repeat protein, PRL1 <em>in vivo</em>. Cell-free protein degradation assay in the absence and presence of proteasome inhibitor indicates that MYB4 is degraded in a ubiquitin proteasome-dependent manner. Furthermore, analyses of MYB4 protein accumulation levels in transgenic <em>atmyb4–1</em> mutant line expressing <em>35</em> <em>S:AtMYB4</em> (<em>35</em> <em>S:AtMYB4-atmyb4–1</em>) and <em>atprl1–1</em> mutant line indicate that PRL1 regulate stability of MYB4 in <em>Arabidopsis</em>. Overall, our results provide important information on the possible mechanism of post-translational modification and regulation of stability of MYB4 protein in <em>Arabidopsis in vivo</em>.</p></div>\",\"PeriodicalId\":38090,\"journal\":{\"name\":\"Current Plant Biology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2024-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2214662824000239/pdfft?md5=2e9b5c0a08002a2e3ecba581f77817ca&pid=1-s2.0-S2214662824000239-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Plant Biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214662824000239\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Plant Biology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214662824000239","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
A nuclear WD40 repeat protein PRL1 regulates stability of MYB4 transcription factor in Arabidopsis
MYB4, a member of the R2R3-type subfamily of MYB transcription factor plays a crucial role in regulating the accumulation of UV-B absorbing phenylpropanoids in plants. UV-B exposure for a longer duration down-regulates the expression of MYB4 gene in Arabidopsis. MYB4 protein represses its own expression by binding to its own promoter. However, at present practically nothing is known about the post-translational regulation of MYB4 protein in vivo. Here, we provide evidence that in Arabidopsis MYB4 protein is phosphorylated in vivo and is targeted by the ubiquitin-26S proteasome-dependent pathway. Immunoprecipitation, immunoblotting, and phosphoprotein staining experiments have revealed that both the accumulation pattern and phosphorylation of MYB4 increase in the light condition during the 24 hours time span under long-day conditions. Yeast two-hybrid and bimolecular fluorescence complementation assays have shown that MYB4 directly interacts with a nuclear WD40 repeat protein, PRL1 in vivo. Cell-free protein degradation assay in the absence and presence of proteasome inhibitor indicates that MYB4 is degraded in a ubiquitin proteasome-dependent manner. Furthermore, analyses of MYB4 protein accumulation levels in transgenic atmyb4–1 mutant line expressing 35S:AtMYB4 (35S:AtMYB4-atmyb4–1) and atprl1–1 mutant line indicate that PRL1 regulate stability of MYB4 in Arabidopsis. Overall, our results provide important information on the possible mechanism of post-translational modification and regulation of stability of MYB4 protein in Arabidopsis in vivo.
期刊介绍:
Current Plant Biology aims to acknowledge and encourage interdisciplinary research in fundamental plant sciences with scope to address crop improvement, biodiversity, nutrition and human health. It publishes review articles, original research papers, method papers and short articles in plant research fields, such as systems biology, cell biology, genetics, epigenetics, mathematical modeling, signal transduction, plant-microbe interactions, synthetic biology, developmental biology, biochemistry, molecular biology, physiology, biotechnologies, bioinformatics and plant genomic resources.