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更正："maea通过β-catenin降解，在爪蟾早期发育过程中影响头部形成"。

IF 1.7 4区生物学 Q4 CELL BIOLOGY

Development Growth & Differentiation Pub Date : 2024-03-18 DOI:10.1111/dgd.12920

{"title":"更正：\"maea通过β-catenin降解，在爪蟾早期发育过程中影响头部形成\"。","authors":"","doi":"10.1111/dgd.12920","DOIUrl":null,"url":null,"abstract":"Goto, T., & Shibuya, H. (2023). maea affects head formation through β-catenin degradation during early Xenopus laevis development. Development, Growth & Differentiation, 65(1), 29–36. https://doi.org/10.1111/dgd.12828In this article, the German letter “Eszett: ß” was used where the Greek letter “beta: β” should have been used in all cases.The following points need to be corrected:In the title,“maea affects head formation through β-catenin degradation during early Xenopus laevis development”In the Abstract,“β-Catenin protein stability is a key factor in canonical Wnt signaling.”“Several E3 ubiquitin ligases contribute to β-catenin degradation through the ubiquitin/proteasome system.”“The expression levels of the Wnt target genes nodal homolog 3, gene 1 (nodal3.1), and siamois homeodomain 1 (sia1), which were induced by injection with β-catenin mRNA, were reduced by maea.S mRNA co-injection. maea.S overexpression at the anterior dorsal region enlarged head structures, whereas Maea knockdown interfered with head formation in Xenopus embryos.”“Maea.S decreased and ubiquitinated β-catenin protein.”“β-catenin-4KRs protein, which mutated the four lysine (K) residues known as ubiquitinated sites to arginine (R) residues, was also ubiquitinated and degraded by Maea.S.”In the KEYWORDS,“degradation, maea, β-catenin, ubiquitination, Xenopus laevis”In the INTRODUCTION (first paragraph),“The key aspect of Wnt signalling is β-catenin protein stability. Disheveled segment polarity protein (Dvl) is recruited at the cell membrane and prevents β-catenin degradation under the Wnt-on state.”“Under the Wnt-off state, Axin1, adenomatous polyposis coli (Apc), casein kinase 1 alpha 1 (Csnk1α1), and glycogen synthase kinase 3 beta (Gsk3β) form the destruction complex to phosphorylate β-catenin protein (Liu et al., 2002).”“Phosphorylated β-catenin is ubiquitinated by E3 ubiquitin ligases, such as beta-transducin repeat-containing E3 ubiquitin-protein ligase (Btrc), and is then degraded by the proteasome system.”In the INTRODUCTION (third paragraph),“There are four lysine residues known as ubiquitinated sites in β-catenin protein. Both lysine residues 19 and 49 are ubiquitinated by Btrc (Winer et al., 2006) and jade family PHD finger 1 (Jade1) (Chitalia et al., 2008).”“Additionally, Siah E3 ubiquitin-protein ligase 1 (Siah1) ubiquitinates β-catenin at lysine residues 666 and 671 (Dimitrova et al., 2010).”“HECT, UBA, and WWE domain containing E3 ubiquitin protein ligase 1 (Huwe1) and SNF2 histone linker PHD RING helicase, E3 ubiquitin protein ligase (Shprh) are also related to β-catenin protein degradation, but the sites they ubiquitinate have not been identified (Dominguez-Brauer et al., 2017; Qu et al., 2016).”In the INTRODUCTION (fourth paragraph),“The Glucose-Induced degradation Deficient (GID) complex also contributes to β-catenin ubiquitination and destruction.”“WD repeat domain 26 (Wdr26), a scaffold protein in the GID complex, degrades β-catenin by binding to Axin1 and is required for head formation in Xenopus (Goto et al., 2016).”“Both human Maea and Rmnd5a ubiquitinate human β-catenin, and their knockdown increases human β-catenin stability in HEK 293 T cells (Sato et al., 2020).”“Therefore, Xenopus Maea could also play an important role in β-catenin degradation in Xenopus.”“However, whether β-catenin protein degradation by Maea occurs in and affects Xenopus development remains unknown.”“Here, we investigated the effects of Xenopus Maea on early Xenopus development through β-catenin degradation.”In the MATERIALS AND METHODS (Section 2.1),“The following primers were used to create a β-catenin-4KRs construct with four lysine (K) to arginine (R) mutations at lysine residues 19, 49, 666, and 671 in the β-catenin amino acid sequence:…”In the MATERIALS AND METHODS (Section 2.2),“Additionally, the following primer pair was used for RT-PCR analysis of β-catenin expression:…”In the MATERIALS AND METHODS (Section 2.5),“For RT-PCR analysis and observing the phenotypes of the injected embryos, we microinjected with maea.S mRNA (250, 500, or 1000 pg/blastomere), β-catenin mRNA (25 or 50 pg/blastomere), β-catenin-4KRs mRNA (25 or 50 pg/blastomere),…”“We microinjected maea.S-FLAG mRNA (500 pg/blastomere) and MYC-β-catenin mRNA (50 pg/blastomere) into four animal blastomeres of 8-cell embryos to obtain lysates.”In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),“Maea.S degraded and ubiquitinated β-catenin”“Knockdown of human Maea suppressed human β-catenin protein degradation (Sato et al., 2020); therefore, we hypothesised that Xenopus maea.S might also regulate the amount of β-catenin protein in Xenopus.”In the figure legend of Figure 2,“β-catenin degradation and ubiquitination by Maea.S.”“(a) WB of ectopically expressed Xenopus β-catenin protein in HEK 293 T cells.”“(b) WB of lysates of Xenopus animal cap cells from embryos (st. 9) injected with MYC-β-catenin mRNA (50 pg/blastomere).”“(c) Cycloheximide chase assay. maea.S-FLAG plasmid was transfected at 24 h after MYC-β-catenin plasmid transfection in HEK 293T cells.”“(d) Interaction between ectopically expressed Xenopus maea.S and β-catenin in HEK 293T cells.”“(e) WB of immunoprecipitates of ubiquitinated β-catenin protein treated with MG-132 (10 μM, 4 h) 24 h after transfection in HEK293T cells.”“(g) RT-PCR analysis of the animal caps from embryos injected with β-catenin mRNA (50 pg/blastomere).”In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),“Therefore, we investigated whether Xenopus maea.S overexpression changed the amount of Xenopus β-catenin protein.”“In HEK 293T cells, transfection of maea.S plasmid reduced the amount of ectopically expressed β-catenin protein in a dose-dependent manner (Figure 2a).”“Additionally, in Xenopus animal caps, ectopically expressed β-catenin protein decreased after injection with maea.S mRNA (Figure 2b).”“Furthermore, the cycloheximide chase assay showed that the transfection of maea.S plasmid promoted β-catenin protein degradation in the short term (4 h) (Figure 2c).”“Moreover, the immunoprecipitation assay revealed that Maea.S also bound to and ubiquitinated β-catenin similar to human proteins, as previously reported (Sato et al., 2020) (Figure 2d,e).”“These results suggest that the degradation system of β-catenin protein by Maea might be similar between humans and Xenopus.”In the RESULTS AND DISCUSSION (Section 3.2, second paragraph),“Because alteration of the transcriptional level of β-catenin by maea was not previously investigated (Sato et al., 2020), we used RT-PCR to confirm β-catenin transcript expression in animal caps of the maea.S mRNA-injected embryos.”“As a result, maea.S mRNA overexpression slightly increased β-catenin transcripts (Figure 2f).”“This suggests that the decrease in β-catenin protein by Maea.S does not occur at the transcriptional level, and the reduction of β-catenin protein by Maea.S upregulates β-catenin transcription to compensate for its protein reduction.”“Moreover, β-catenin mRNA overexpression in animal caps slightly increased maea expression (Figure 2g).”“In Xenopus, there may be a system for mutually controlling transcript levels to maintain a steady level of β-catenin protein.”In the RESULTS AND DISCUSSION (Section 3.3, title),“Overexpression of maea.S mRNA inhibited the effects of β-catenin”In the RESULTS AND DISCUSSION (Section 3.3, first paragraph),“To confirm the effect of β-catenin protein degradation by Maea.S in Xenopus development,…”In the RESULTS AND DISCUSSION (Section 3.3, second paragraph),“Because anterior Wnt inhibition is necessary for head formation (De Robertis & Kuroda, 2004; Ding et al., 2018; Kumar et al., 2021; Niehrs, 2022), the enlargement of head structures of embryos injected with maea.S mRNA might be caused by β-catenin protein degradation in the anterior region.”“Therefore, Rmnd5a might also contribute to β-catenin protein degradation by working with Maea in Xenopus in a similar manner to previously reported human cultured cells (Sato et al., 2020).”In the RESULTS AND DISCUSSION (Section 3.3, third paragraph),“When we injected a low dose of β-catenin mRNA into ventral blastomeres of 4-cell embryos,…”“Wnt target gene expression in embryos ventrally injected with β-catenin mRNA also decreased by co-injection with maea.S mRNA (Figure 3e).”“These results suggest that maea.S may function as a gene that suppresses excessive Wnt activities through the degradation of β-catenin protein during early development.”In the RESULTS AND DISCUSSION (Section 3.4, first paragraph),“These findings reveal that maea might contribute to head formation by inhibiting the Wnt activity through β-catenin protein degradation during early embryogenesis.”In the figure legend of Figure 3,“Effects of β-catenin on head and secondary axis formation by maea.S.”“(c) Phenotypes of embryos (st. 30) when injected with β-catenin mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage.”“(e) RT-PCR analysis of the ventral sectors from embryos injected with β-catenin mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”In the figure legend of Figure 4,“Inhibition effects of maea.S on the β-catenin-4KRs construct.”“(a) WB of ectopically expressed β-catenin-4KRs in HEK 293T cells.”“(b) Interaction between ectopically expressed Maea.S and β-catenin- 4KRs in HEK 293T cells.”“(c) WB of ubiquitinated β-catenin-4KRs treated as described in Figure 2e.”“(d) The appearance rates of embryo phenotypes when injected with β-catenin-4KRs mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage (see Figure 3c,d).”“(e) RT-PCR analysis of the ventral sectors of embryos injected with β-catenin-4KRs mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”In the RESULTS AND DISCUSSION (Section 3.5, title),“Maea.S might ubiquitinate unknown lysine residues of β-catenin”In the RESULTS AND DISCUSSION (Section 3.5, first paragraph),“In vertebrates, the amino acid sequence of β-catenin involves 26 lysine residues, which are conserved amongst species.”“In the GID complex proteins, Wdr26 and Rmnd5a also seem to contribute to β-catenin degradation…”In the RESULTS AND DISCUSSION (Section 3.5, second paragraph),“To investigate whether Maea.S ubiquitinates unknown lysine residues of β-catenin protein, we used a β-catenin-4KRs construct with four lysine to arginine mutations at 19, 49, 666, and 671 lysine residues.”“In HEK 293T cells, maea.S plasmid transfection reduced β-catenin-4KRs protein amounts (Figure 4a).”“Moreover, Maea.S also bound to and ubiquitinated β-catenin-4KRs protein (Figure 4b,c).”“The appearance rates of the secondary axis with complete and partial head structures of embryos injected with β-catenin-4KRs mRNA were decreased by maea.S mRNA co-injection (Figure 4d).”“Wnt target gene expression of embryos ventrally injected with β-catenin-4KRs mRNA was also reduced by co-injection with maea.S mRNA (Figure 4e).”“These results demonstrate that β-catenin protein degradation by Maea.S might be due to β-catenin protein ubiquitination at unknown lysine residues.”“However, because degradation of β-catenin-4KRs protein by Maea.S was less efficient than that of β-catenin protein, Maea.S might also ubiquitinate parts of or all known lysine residues (Figures 2a and 4a).”The online article has been corrected.We apologize for this error.","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"66 3","pages":"266-270"},"PeriodicalIF":1.7000,"publicationDate":"2024-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/dgd.12920","citationCount":"0","resultStr":"{\"title\":\"Correction to “maea affects head formation through β-catenin degradation during early Xenopus laevis development”\",\"authors\":\"\",\"doi\":\"10.1111/dgd.12920\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Goto, T., & Shibuya, H. (2023). maea affects head formation through β-catenin degradation during early Xenopus laevis development. Development, Growth & Differentiation, 65(1), 29–36. https://doi.org/10.1111/dgd.12828In this article, the German letter “Eszett: ß” was used where the Greek letter “beta: β” should have been used in all cases.The following points need to be corrected:In the title,“maea affects head formation through β-catenin degradation during early Xenopus laevis development”In the Abstract,“β-Catenin protein stability is a key factor in canonical Wnt signaling.”“Several E3 ubiquitin ligases contribute to β-catenin degradation through the ubiquitin/proteasome system.”“The expression levels of the Wnt target genes nodal homolog 3, gene 1 (nodal3.1), and siamois homeodomain 1 (sia1), which were induced by injection with β-catenin mRNA, were reduced by maea.S mRNA co-injection. maea.S overexpression at the anterior dorsal region enlarged head structures, whereas Maea knockdown interfered with head formation in Xenopus embryos.”“Maea.S decreased and ubiquitinated β-catenin protein.”“β-catenin-4KRs protein, which mutated the four lysine (K) residues known as ubiquitinated sites to arginine (R) residues, was also ubiquitinated and degraded by Maea.S.”In the KEYWORDS,“degradation, maea, β-catenin, ubiquitination, Xenopus laevis”In the INTRODUCTION (first paragraph),“The key aspect of Wnt signalling is β-catenin protein stability. Disheveled segment polarity protein (Dvl) is recruited at the cell membrane and prevents β-catenin degradation under the Wnt-on state.”“Under the Wnt-off state, Axin1, adenomatous polyposis coli (Apc), casein kinase 1 alpha 1 (Csnk1α1), and glycogen synthase kinase 3 beta (Gsk3β) form the destruction complex to phosphorylate β-catenin protein (Liu et al., 2002).”“Phosphorylated β-catenin is ubiquitinated by E3 ubiquitin ligases, such as beta-transducin repeat-containing E3 ubiquitin-protein ligase (Btrc), and is then degraded by the proteasome system.”In the INTRODUCTION (third paragraph),“There are four lysine residues known as ubiquitinated sites in β-catenin protein. Both lysine residues 19 and 49 are ubiquitinated by Btrc (Winer et al., 2006) and jade family PHD finger 1 (Jade1) (Chitalia et al., 2008).”“Additionally, Siah E3 ubiquitin-protein ligase 1 (Siah1) ubiquitinates β-catenin at lysine residues 666 and 671 (Dimitrova et al., 2010).”“HECT, UBA, and WWE domain containing E3 ubiquitin protein ligase 1 (Huwe1) and SNF2 histone linker PHD RING helicase, E3 ubiquitin protein ligase (Shprh) are also related to β-catenin protein degradation, but the sites they ubiquitinate have not been identified (Dominguez-Brauer et al., 2017; Qu et al., 2016).”In the INTRODUCTION (fourth paragraph),“The Glucose-Induced degradation Deficient (GID) complex also contributes to β-catenin ubiquitination and destruction.”“WD repeat domain 26 (Wdr26), a scaffold protein in the GID complex, degrades β-catenin by binding to Axin1 and is required for head formation in Xenopus (Goto et al., 2016).”“Both human Maea and Rmnd5a ubiquitinate human β-catenin, and their knockdown increases human β-catenin stability in HEK 293 T cells (Sato et al., 2020).”“Therefore, Xenopus Maea could also play an important role in β-catenin degradation in Xenopus.”“However, whether β-catenin protein degradation by Maea occurs in and affects Xenopus development remains unknown.”“Here, we investigated the effects of Xenopus Maea on early Xenopus development through β-catenin degradation.”In the MATERIALS AND METHODS (Section 2.1),“The following primers were used to create a β-catenin-4KRs construct with four lysine (K) to arginine (R) mutations at lysine residues 19, 49, 666, and 671 in the β-catenin amino acid sequence:…”In the MATERIALS AND METHODS (Section 2.2),“Additionally, the following primer pair was used for RT-PCR analysis of β-catenin expression:…”In the MATERIALS AND METHODS (Section 2.5),“For RT-PCR analysis and observing the phenotypes of the injected embryos, we microinjected with maea.S mRNA (250, 500, or 1000 pg/blastomere), β-catenin mRNA (25 or 50 pg/blastomere), β-catenin-4KRs mRNA (25 or 50 pg/blastomere),…”“We microinjected maea.S-FLAG mRNA (500 pg/blastomere) and MYC-β-catenin mRNA (50 pg/blastomere) into four animal blastomeres of 8-cell embryos to obtain lysates.”In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),“Maea.S degraded and ubiquitinated β-catenin”“Knockdown of human Maea suppressed human β-catenin protein degradation (Sato et al., 2020); therefore, we hypothesised that Xenopus maea.S might also regulate the amount of β-catenin protein in Xenopus.”In the figure legend of Figure 2,“β-catenin degradation and ubiquitination by Maea.S.”“(a) WB of ectopically expressed Xenopus β-catenin protein in HEK 293 T cells.”“(b) WB of lysates of Xenopus animal cap cells from embryos (st. 9) injected with MYC-β-catenin mRNA (50 pg/blastomere).”“(c) Cycloheximide chase assay. maea.S-FLAG plasmid was transfected at 24 h after MYC-β-catenin plasmid transfection in HEK 293T cells.”“(d) Interaction between ectopically expressed Xenopus maea.S and β-catenin in HEK 293T cells.”“(e) WB of immunoprecipitates of ubiquitinated β-catenin protein treated with MG-132 (10 μM, 4 h) 24 h after transfection in HEK293T cells.”“(g) RT-PCR analysis of the animal caps from embryos injected with β-catenin mRNA (50 pg/blastomere).”In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),“Therefore, we investigated whether Xenopus maea.S overexpression changed the amount of Xenopus β-catenin protein.”“In HEK 293T cells, transfection of maea.S plasmid reduced the amount of ectopically expressed β-catenin protein in a dose-dependent manner (Figure 2a).”“Additionally, in Xenopus animal caps, ectopically expressed β-catenin protein decreased after injection with maea.S mRNA (Figure 2b).”“Furthermore, the cycloheximide chase assay showed that the transfection of maea.S plasmid promoted β-catenin protein degradation in the short term (4 h) (Figure 2c).”“Moreover, the immunoprecipitation assay revealed that Maea.S also bound to and ubiquitinated β-catenin similar to human proteins, as previously reported (Sato et al., 2020) (Figure 2d,e).”“These results suggest that the degradation system of β-catenin protein by Maea might be similar between humans and Xenopus.”In the RESULTS AND DISCUSSION (Section 3.2, second paragraph),“Because alteration of the transcriptional level of β-catenin by maea was not previously investigated (Sato et al., 2020), we used RT-PCR to confirm β-catenin transcript expression in animal caps of the maea.S mRNA-injected embryos.”“As a result, maea.S mRNA overexpression slightly increased β-catenin transcripts (Figure 2f).”“This suggests that the decrease in β-catenin protein by Maea.S does not occur at the transcriptional level, and the reduction of β-catenin protein by Maea.S upregulates β-catenin transcription to compensate for its protein reduction.”“Moreover, β-catenin mRNA overexpression in animal caps slightly increased maea expression (Figure 2g).”“In Xenopus, there may be a system for mutually controlling transcript levels to maintain a steady level of β-catenin protein.”In the RESULTS AND DISCUSSION (Section 3.3, title),“Overexpression of maea.S mRNA inhibited the effects of β-catenin”In the RESULTS AND DISCUSSION (Section 3.3, first paragraph),“To confirm the effect of β-catenin protein degradation by Maea.S in Xenopus development,…”In the RESULTS AND DISCUSSION (Section 3.3, second paragraph),“Because anterior Wnt inhibition is necessary for head formation (De Robertis & Kuroda, 2004; Ding et al., 2018; Kumar et al., 2021; Niehrs, 2022), the enlargement of head structures of embryos injected with maea.S mRNA might be caused by β-catenin protein degradation in the anterior region.”“Therefore, Rmnd5a might also contribute to β-catenin protein degradation by working with Maea in Xenopus in a similar manner to previously reported human cultured cells (Sato et al., 2020).”In the RESULTS AND DISCUSSION (Section 3.3, third paragraph),“When we injected a low dose of β-catenin mRNA into ventral blastomeres of 4-cell embryos,…”“Wnt target gene expression in embryos ventrally injected with β-catenin mRNA also decreased by co-injection with maea.S mRNA (Figure 3e).”“These results suggest that maea.S may function as a gene that suppresses excessive Wnt activities through the degradation of β-catenin protein during early development.”In the RESULTS AND DISCUSSION (Section 3.4, first paragraph),“These findings reveal that maea might contribute to head formation by inhibiting the Wnt activity through β-catenin protein degradation during early embryogenesis.”In the figure legend of Figure 3,“Effects of β-catenin on head and secondary axis formation by maea.S.”“(c) Phenotypes of embryos (st. 30) when injected with β-catenin mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage.”“(e) RT-PCR analysis of the ventral sectors from embryos injected with β-catenin mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”In the figure legend of Figure 4,“Inhibition effects of maea.S on the β-catenin-4KRs construct.”“(a) WB of ectopically expressed β-catenin-4KRs in HEK 293T cells.”“(b) Interaction between ectopically expressed Maea.S and β-catenin- 4KRs in HEK 293T cells.”“(c) WB of ubiquitinated β-catenin-4KRs treated as described in Figure 2e.”“(d) The appearance rates of embryo phenotypes when injected with β-catenin-4KRs mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage (see Figure 3c,d).”“(e) RT-PCR analysis of the ventral sectors of embryos injected with β-catenin-4KRs mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”In the RESULTS AND DISCUSSION (Section 3.5, title),“Maea.S might ubiquitinate unknown lysine residues of β-catenin”In the RESULTS AND DISCUSSION (Section 3.5, first paragraph),“In vertebrates, the amino acid sequence of β-catenin involves 26 lysine residues, which are conserved amongst species.”“In the GID complex proteins, Wdr26 and Rmnd5a also seem to contribute to β-catenin degradation…”In the RESULTS AND DISCUSSION (Section 3.5, second paragraph),“To investigate whether Maea.S ubiquitinates unknown lysine residues of β-catenin protein, we used a β-catenin-4KRs construct with four lysine to arginine mutations at 19, 49, 666, and 671 lysine residues.”“In HEK 293T cells, maea.S plasmid transfection reduced β-catenin-4KRs protein amounts (Figure 4a).”“Moreover, Maea.S also bound to and ubiquitinated β-catenin-4KRs protein (Figure 4b,c).”“The appearance rates of the secondary axis with complete and partial head structures of embryos injected with β-catenin-4KRs mRNA were decreased by maea.S mRNA co-injection (Figure 4d).”“Wnt target gene expression of embryos ventrally injected with β-catenin-4KRs mRNA was also reduced by co-injection with maea.S mRNA (Figure 4e).”“These results demonstrate that β-catenin protein degradation by Maea.S might be due to β-catenin protein ubiquitination at unknown lysine residues.”“However, because degradation of β-catenin-4KRs protein by Maea.S was less efficient than that of β-catenin protein, Maea.S might also ubiquitinate parts of or all known lysine residues (Figures 2a and 4a).”The online article has been corrected.We apologize for this error.\",\"PeriodicalId\":50589,\"journal\":{\"name\":\"Development Growth & Differentiation\",\"volume\":\"66 3\",\"pages\":\"266-270\"},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2024-03-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/dgd.12920\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Development Growth & Differentiation\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/dgd.12920\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Development Growth & Differentiation","FirstCategoryId":"99","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/dgd.12920","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}

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摘要

""(d)异位表达的 Xenopus maea.S 和 β-catenin 在 HEK 293T 细胞中的相互作用。""(e)转染 HEK293T 细胞 24 h 后，用 MG-132（10 μM，4 h）处理泛素化的β-catenin 蛋白的免疫沉淀 WB。"结果与讨论（第 3.2 节，第一段）中说："因此，我们研究了 Xenopus maea.S 的过表达是否会改变 Xenopus β-catenin 蛋白的量。""在 HEK 293T 细胞中，转染 maea.S 质粒后，异位表达的β-catenin 蛋白量减少，且呈剂量依赖性（图 2a）。""此外，在爪蟾动物巢穴中，注射 maea.S mRNA 后，异位表达的β-catenin 蛋白量减少（图 2b）。""此外，环己亚胺追逐试验显示，转染 maea.S 质粒可在短期内（4 小时）促进 β-catenin 蛋白降解（图 2c）。"""而且，免疫沉淀试验显示，Maea.S 也与人类蛋白类似，与 β-catenin 结合并泛素化（Sato et al、2020）（图 2d，e）。""这些结果表明，Maea 对 β-catenin 蛋白的降解系统可能与人类和爪蟾相似。"在 "结果与讨论"（第 3.2 节，第二段）中，"由于以前没有研究过 maea 对 β-catenin 转录水平的改变（Sato et al、结果，maea.S mRNA 的过表达略微增加了 β-catenin转录本（图 2f）""这表明，Maea.S 对 β-catenin蛋白的减少并不是发生在转录水平上。""这表明，Maea.S 对 β-catenin蛋白的减少并不是在转录水平上发生的，Maea.S 对 β-catenin蛋白的减少会上调 β-catenin的转录以补偿其蛋白的减少。""此外，在动物盖中过量表达 β-cateninmRNA会略微增加 maea 的表达（图 2g）。"在结果与讨论（第 3.3 节，标题）中，"maea.S mRNA 的过表达抑制了 β-catenin 的作用 "在结果与讨论（第 3.3节，第一段）中，"为了证实Maea.S在章鱼发育过程中降解β-catenin蛋白的作用，...... "在结果与讨论（第3.3节，第二段）中，"因为前Wnt抑制是头部形成的必要条件（De Robertis & Kuroda, 2004; Ding et al、2018；Kumar et al.，2021；Niehrs，2022），注射了 maea.S mRNA 的胚胎头部结构的增大可能是由前部区域的 β-catenin 蛋白降解引起的。""因此，Rmnd5a 也可能通过在爪蟾中与 Maea 合作，以类似于之前报道的人类培养细胞的方式促进 β-catenin 蛋白降解（Sato et al、在 "结果与讨论"（第 3.3 节，第三段）中，"当我们将低剂量的 β-catenin mRNA 注入 4 细胞胚胎的腹腔胚泡时，......""在腹腔注入 β-catenin mRNA 的胚胎中，Wnt 靶基因的表达也因同时注入 maea.这些结果表明，maea.S 的功能可能是在早期发育过程中通过降解 β-catenin 蛋白来抑制过度的 Wnt 活性。"在结果与讨论（第 3.4 节，第一段）中，"这些发现揭示了 maea 可能在早期胚胎发育过程中通过降解 β-catenin 蛋白来抑制 Wnt 活性，从而促进头部的形成。"在图 3 的图例中，"β-catenin 对 maea.S 的头部和次轴形成的影响""(c) 胚胎（st. 30）在 4 细胞期向腹侧胚泡注射 β-catenin mRNA 或与 maea.S mRNA 联合注射时的表型。"图 4 的图例中，"maea.S 对 β-catenin-4KRs 构建体的抑制作用"。""(a)在 HEK 293T 细胞中异位表达的 β-catenin-4KRs 的 WB。""(b)在 HEK 293T 细胞中异位表达的 Maea.S 与 β-catenin- 4KRs 之间的相互作用。""(c)如图 2e 所述处理泛素化的β-catenin-4KRs 的 WB。""(d)在 4 细胞期向腹侧胚泡注射 β-catenin-4KRs mRNA 或与 maea.S mRNA 联合注射时胚胎表型的出现率（见图 3c，d）。""(e) RT-PCR 分析向腹侧胚泡注射 β-catenin-4KRs mRNA（50 pg/胚泡）或与 maea.S mRNA 联合注射时胚胎表型的出现率（见图 3d）。在结果与讨论（第 3.5 节，标题）中，"Maea.S 可能泛素化 β-catenin 的未知赖氨酸残基"。

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Correction to “maea affects head formation through β-catenin degradation during early Xenopus laevis development”

Goto, T., & Shibuya, H. (2023). maea affects head formation through β-catenin degradation during early Xenopus laevis development. Development, Growth & Differentiation, 65(1), 29–36. https://doi.org/10.1111/dgd.12828

In this article, the German letter “Eszett: ß” was used where the Greek letter “beta: β” should have been used in all cases.

The following points need to be corrected:

In the title,

“maea affects head formation through β-catenin degradation during early Xenopus laevis development”

In the Abstract,

“β-Catenin protein stability is a key factor in canonical Wnt signaling.”

“Several E3 ubiquitin ligases contribute to β-catenin degradation through the ubiquitin/proteasome system.”

“The expression levels of the Wnt target genes nodal homolog 3, gene 1 (nodal3.1), and siamois homeodomain 1 (sia1), which were induced by injection with β-catenin mRNA, were reduced by maea.S mRNA co-injection. maea.S overexpression at the anterior dorsal region enlarged head structures, whereas Maea knockdown interfered with head formation in Xenopus embryos.”

“Maea.S decreased and ubiquitinated β-catenin protein.”

“β-catenin-4KRs protein, which mutated the four lysine (K) residues known as ubiquitinated sites to arginine (R) residues, was also ubiquitinated and degraded by Maea.S.”

In the KEYWORDS,

“degradation, maea, β-catenin, ubiquitination, Xenopus laevis”

In the INTRODUCTION (first paragraph),

“The key aspect of Wnt signalling is β-catenin protein stability. Disheveled segment polarity protein (Dvl) is recruited at the cell membrane and prevents β-catenin degradation under the Wnt-on state.”

“Under the Wnt-off state, Axin1, adenomatous polyposis coli (Apc), casein kinase 1 alpha 1 (Csnk1α1), and glycogen synthase kinase 3 beta (Gsk3β) form the destruction complex to phosphorylate β-catenin protein (Liu et al., 2002).”

“Phosphorylated β-catenin is ubiquitinated by E3 ubiquitin ligases, such as beta-transducin repeat-containing E3 ubiquitin-protein ligase (Btrc), and is then degraded by the proteasome system.”

In the INTRODUCTION (third paragraph),

“There are four lysine residues known as ubiquitinated sites in β-catenin protein. Both lysine residues 19 and 49 are ubiquitinated by Btrc (Winer et al., 2006) and jade family PHD finger 1 (Jade1) (Chitalia et al., 2008).”

“Additionally, Siah E3 ubiquitin-protein ligase 1 (Siah1) ubiquitinates β-catenin at lysine residues 666 and 671 (Dimitrova et al., 2010).”

“HECT, UBA, and WWE domain containing E3 ubiquitin protein ligase 1 (Huwe1) and SNF2 histone linker PHD RING helicase, E3 ubiquitin protein ligase (Shprh) are also related to β-catenin protein degradation, but the sites they ubiquitinate have not been identified (Dominguez-Brauer et al., 2017; Qu et al., 2016).”

In the INTRODUCTION (fourth paragraph),

“The Glucose-Induced degradation Deficient (GID) complex also contributes to β-catenin ubiquitination and destruction.”

“WD repeat domain 26 (Wdr26), a scaffold protein in the GID complex, degrades β-catenin by binding to Axin1 and is required for head formation in Xenopus (Goto et al., 2016).”

“Both human Maea and Rmnd5a ubiquitinate human β-catenin, and their knockdown increases human β-catenin stability in HEK 293 T cells (Sato et al., 2020).”

“Therefore, Xenopus Maea could also play an important role in β-catenin degradation in Xenopus.”

“However, whether β-catenin protein degradation by Maea occurs in and affects Xenopus development remains unknown.”

“Here, we investigated the effects of Xenopus Maea on early Xenopus development through β-catenin degradation.”

In the MATERIALS AND METHODS (Section 2.1),

“The following primers were used to create a β-catenin-4KRs construct with four lysine (K) to arginine (R) mutations at lysine residues 19, 49, 666, and 671 in the β-catenin amino acid sequence:…”

In the MATERIALS AND METHODS (Section 2.2),

“Additionally, the following primer pair was used for RT-PCR analysis of β-catenin expression:…”

In the MATERIALS AND METHODS (Section 2.5),

“For RT-PCR analysis and observing the phenotypes of the injected embryos, we microinjected with maea.S mRNA (250, 500, or 1000 pg/blastomere), β-catenin mRNA (25 or 50 pg/blastomere), β-catenin-4KRs mRNA (25 or 50 pg/blastomere),…”

“We microinjected maea.S-FLAG mRNA (500 pg/blastomere) and MYC-β-catenin mRNA (50 pg/blastomere) into four animal blastomeres of 8-cell embryos to obtain lysates.”

In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),

“Maea.S degraded and ubiquitinated β-catenin”

“Knockdown of human Maea suppressed human β-catenin protein degradation (Sato et al., 2020); therefore, we hypothesised that Xenopus maea.S might also regulate the amount of β-catenin protein in Xenopus.”

In the figure legend of Figure 2,

“β-catenin degradation and ubiquitination by Maea.S.”

“(a) WB of ectopically expressed Xenopus β-catenin protein in HEK 293 T cells.”

“(b) WB of lysates of Xenopus animal cap cells from embryos (st. 9) injected with MYC-β-catenin mRNA (50 pg/blastomere).”

“(c) Cycloheximide chase assay. maea.S-FLAG plasmid was transfected at 24 h after MYC-β-catenin plasmid transfection in HEK 293T cells.”

“(d) Interaction between ectopically expressed Xenopus maea.S and β-catenin in HEK 293T cells.”

“(e) WB of immunoprecipitates of ubiquitinated β-catenin protein treated with MG-132 (10 μM, 4 h) 24 h after transfection in HEK293T cells.”

“(g) RT-PCR analysis of the animal caps from embryos injected with β-catenin mRNA (50 pg/blastomere).”

In the RESULTS AND DISCUSSION (Section 3.2, first paragraph),

“Therefore, we investigated whether Xenopus maea.S overexpression changed the amount of Xenopus β-catenin protein.”

“In HEK 293T cells, transfection of maea.S plasmid reduced the amount of ectopically expressed β-catenin protein in a dose-dependent manner (Figure 2a).”

“Additionally, in Xenopus animal caps, ectopically expressed β-catenin protein decreased after injection with maea.S mRNA (Figure 2b).”

“Furthermore, the cycloheximide chase assay showed that the transfection of maea.S plasmid promoted β-catenin protein degradation in the short term (4 h) (Figure 2c).”

“Moreover, the immunoprecipitation assay revealed that Maea.S also bound to and ubiquitinated β-catenin similar to human proteins, as previously reported (Sato et al., 2020) (Figure 2d,e).”

“These results suggest that the degradation system of β-catenin protein by Maea might be similar between humans and Xenopus.”

In the RESULTS AND DISCUSSION (Section 3.2, second paragraph),

“Because alteration of the transcriptional level of β-catenin by maea was not previously investigated (Sato et al., 2020), we used RT-PCR to confirm β-catenin transcript expression in animal caps of the maea.S mRNA-injected embryos.”

“As a result, maea.S mRNA overexpression slightly increased β-catenin transcripts (Figure 2f).”

“This suggests that the decrease in β-catenin protein by Maea.S does not occur at the transcriptional level, and the reduction of β-catenin protein by Maea.S upregulates β-catenin transcription to compensate for its protein reduction.”

“Moreover, β-catenin mRNA overexpression in animal caps slightly increased maea expression (Figure 2g).”

“In Xenopus, there may be a system for mutually controlling transcript levels to maintain a steady level of β-catenin protein.”

In the RESULTS AND DISCUSSION (Section 3.3, title),

“Overexpression of maea.S mRNA inhibited the effects of β-catenin”

In the RESULTS AND DISCUSSION (Section 3.3, first paragraph),

“To confirm the effect of β-catenin protein degradation by Maea.S in Xenopus development,…”

In the RESULTS AND DISCUSSION (Section 3.3, second paragraph),

“Because anterior Wnt inhibition is necessary for head formation (De Robertis & Kuroda, 2004; Ding et al., 2018; Kumar et al., 2021; Niehrs, 2022), the enlargement of head structures of embryos injected with maea.S mRNA might be caused by β-catenin protein degradation in the anterior region.”

“Therefore, Rmnd5a might also contribute to β-catenin protein degradation by working with Maea in Xenopus in a similar manner to previously reported human cultured cells (Sato et al., 2020).”

In the RESULTS AND DISCUSSION (Section 3.3, third paragraph),

“When we injected a low dose of β-catenin mRNA into ventral blastomeres of 4-cell embryos,…”

“Wnt target gene expression in embryos ventrally injected with β-catenin mRNA also decreased by co-injection with maea.S mRNA (Figure 3e).”

“These results suggest that maea.S may function as a gene that suppresses excessive Wnt activities through the degradation of β-catenin protein during early development.”

In the RESULTS AND DISCUSSION (Section 3.4, first paragraph),

“These findings reveal that maea might contribute to head formation by inhibiting the Wnt activity through β-catenin protein degradation during early embryogenesis.”

In the figure legend of Figure 3,

“Effects of β-catenin on head and secondary axis formation by maea.S.”

“(c) Phenotypes of embryos (st. 30) when injected with β-catenin mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage.”

“(e) RT-PCR analysis of the ventral sectors from embryos injected with β-catenin mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”

In the figure legend of Figure 4,

“Inhibition effects of maea.S on the β-catenin-4KRs construct.”

“(a) WB of ectopically expressed β-catenin-4KRs in HEK 293T cells.”

“(b) Interaction between ectopically expressed Maea.S and β-catenin- 4KRs in HEK 293T cells.”

“(c) WB of ubiquitinated β-catenin-4KRs treated as described in Figure 2e.”

“(d) The appearance rates of embryo phenotypes when injected with β-catenin-4KRs mRNA or co-injected with maea.S mRNA into ventral blastomeres at the 4-cell stage (see Figure 3c,d).”

“(e) RT-PCR analysis of the ventral sectors of embryos injected with β-catenin-4KRs mRNA (50 pg/blastomere) or co-injected with maea.S mRNA (500 pg/blastomere).”

In the RESULTS AND DISCUSSION (Section 3.5, title),

“Maea.S might ubiquitinate unknown lysine residues of β-catenin”

In the RESULTS AND DISCUSSION (Section 3.5, first paragraph),

“In vertebrates, the amino acid sequence of β-catenin involves 26 lysine residues, which are conserved amongst species.”

“In the GID complex proteins, Wdr26 and Rmnd5a also seem to contribute to β-catenin degradation…”

In the RESULTS AND DISCUSSION (Section 3.5, second paragraph),

“To investigate whether Maea.S ubiquitinates unknown lysine residues of β-catenin protein, we used a β-catenin-4KRs construct with four lysine to arginine mutations at 19, 49, 666, and 671 lysine residues.”

“In HEK 293T cells, maea.S plasmid transfection reduced β-catenin-4KRs protein amounts (Figure 4a).”

“Moreover, Maea.S also bound to and ubiquitinated β-catenin-4KRs protein (Figure 4b,c).”

“The appearance rates of the secondary axis with complete and partial head structures of embryos injected with β-catenin-4KRs mRNA were decreased by maea.S mRNA co-injection (Figure 4d).”

“Wnt target gene expression of embryos ventrally injected with β-catenin-4KRs mRNA was also reduced by co-injection with maea.S mRNA (Figure 4e).”

“These results demonstrate that β-catenin protein degradation by Maea.S might be due to β-catenin protein ubiquitination at unknown lysine residues.”

“However, because degradation of β-catenin-4KRs protein by Maea.S was less efficient than that of β-catenin protein, Maea.S might also ubiquitinate parts of or all known lysine residues (Figures 2a and 4a).”

The online article has been corrected.

We apologize for this error.

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来源期刊

Development Growth & Differentiation 生物-发育生物学

CiteScore

4.60

自引率

4.00%

发文量

审稿时长

6 months

期刊介绍： Development Growth & Differentiation (DGD) publishes three types of articles: original, resource, and review papers. Original papers are on any subjects having a context in development, growth, and differentiation processes in animals, plants, and microorganisms, dealing with molecular, genetic, cellular and organismal phenomena including metamorphosis and regeneration, while using experimental, theoretical, and bioinformatic approaches. Papers on other related fields are also welcome, such as stem cell biology, genomics, neuroscience, Evodevo, Ecodevo, and medical science as well as related methodology (new or revised techniques) and bioresources. Resource papers describe a dataset, such as whole genome sequences and expressed sequence tags (ESTs), with some biological insights, which should be valuable for studying the subjects as mentioned above. Submission of review papers is also encouraged, especially those providing a new scope based on the authors’ own study, or a summarization of their study series.