Protein biosynthesis by the ribosome is a fundamental biological process in living systems.Recent studies suggest that ribosomal subunits also play essential roles in cell growth and differentiation beyond their roles...Protein biosynthesis by the ribosome is a fundamental biological process in living systems.Recent studies suggest that ribosomal subunits also play essential roles in cell growth and differentiation beyond their roles in protein translation.The ribosomal subunit RPS6 has been studied for more than 50 years in various organisms,but little is known about its specific roles in certain signaling pathways.In this study,we focused on the functions of Arabidopsis RPS6A in auxin-related root growth and development.The rps6a mutant presented a series of auxin-deficient phenotypes,such as shortened primary roots,reduced lateral root numbers,and defective vasculatures.Treatment of the rps6a mutant with various concentrations of auxin and its analogs did not restore the root defect phenotypes,suggesting a defect in the auxin signaling pathway.Further cell biological and global transcriptome analyses revealed that auxin signaling was weakened in the rps6a mutant and that there was a reduced abundance of PIN-FORMED(PIN)auxin transporters.Our work provides insights into the role of the protein biosynthesis pathway involved in auxin signaling.展开更多
Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to ...Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role.Methods Initially,we assessed PES1 expression patterns in two distinct senescence models:replicative senescent mouse embryonic fibroblasts(MEFs)and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells.Subsequently,PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types.Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays,respectively.The expression of senescence-associated proteins(p53,p21,and Rb)and SASP factors(IL-6,IL-1β,and IL-8)were analyzed by Western blot or qPCR.Furthermore,Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology.Results PES1 expression was significantly downregulated in senescent MEFs and HepG2 cells.PES1 knockdown resulted in decreased EdU-positive cells and increased SA-β-gal-positive cells,indicating proliferation inhibition and senescence induction.Mechanistically,PES1 suppression activated the p53-p21 pathway without affecting Rb expression,while upregulating IL-6,IL-1β,and IL-8 production.Notably,PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress,as evidenced by aberrant nucleolar morphology.Conclusion Our findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent(but Rb-independent)cellular senescence,highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.展开更多
背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细...背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细胞(dentritic cell,DC),探讨负载RPL8蛋白的DC对黑色素瘤的免疫效应。方法:原核表达RPL8蛋白,纯化后致敏小鼠骨髓来源DC,流式细胞仪检测DC表面标志,MTT法检测细胞毒性T淋巴细胞对小鼠黑色素瘤细胞的杀伤作用;负载RPL8蛋白DC免疫治疗小鼠后,观察肿瘤体积变化及小鼠生存时间。结果:纯化蛋白经蛋白质印迹法(Western blot)分析见约28×103大小的特异性条带;DC经RPL8及细菌脂多糖(Lipoplysaccharide,LPS)诱导成熟后细胞表面CD11c、CD80、MHC-Ⅰ类、MHC-Ⅱ类分子表达增高,能激活T淋巴细胞,对B16细胞有抑制作用,RPL8-DC组的抑制率在效靶比为30∶1时高达70%,较PBS组和DC组明显增高;负载RPL8蛋白DC免疫治疗小鼠后,肿瘤体积缩小,小鼠的生存期明显延长。结论:负载RPL8的DC对黑色素瘤有生长抑制作用。展开更多
Objective: In order to coordinately express amarker gene and therapeutic genes in one vector, apolycistronic retroviral vector was constructed withthe internal ribosome entry site from encephalo-myocarditis and polio ...Objective: In order to coordinately express amarker gene and therapeutic genes in one vector, apolycistronic retroviral vector was constructed withthe internal ribosome entry site from encephalo-myocarditis and polio virus, which was able to expressthree different genes simultaneously. Methods: Thevector used mouse moloney leukemia virus sequencesfor viral expression and packaging functions展开更多
Interleukin-12, a heterodimer produced mainlyby antigen presenting cells (APCs), plays vital rolesin the induction of Thl-mediated immune responsesand has the potential to treat cancer and infectiousdiseases. Systemic...Interleukin-12, a heterodimer produced mainlyby antigen presenting cells (APCs), plays vital rolesin the induction of Thl-mediated immune responsesand has the potential to treat cancer and infectiousdiseases. Systemic administration of IL-12 wasobserved to be associated with severe toxicity. To takefull advantage of its immunoregulatory activities andminimize its toxicities, local administration of IL-12may be the ideal approach. Intratumoral injection展开更多
Soybean is one of the most important sources of edible oil and proteins in the world. However, it suffers from many kinds of fungal diseases which is a major limiting factor in soybean production. The fungal disease c...Soybean is one of the most important sources of edible oil and proteins in the world. However, it suffers from many kinds of fungal diseases which is a major limiting factor in soybean production. The fungal disease can be effectively controlled by breeding plant cultivars with genetic transformation. In this study, the resistance to Phytophthora sojae of five bivalent transgenic soybean lines was identified using the hypocotyls inoculation technique. The lines were the T2 of the transgenic soybean which were transformed with kidney bean chitinase gene and barley ribosome inactivating protein gene, and were positive by Southern Blot analysis. The resistance difference was studied through comparing the death percentage of transgenic soybean with the control. The results showed that four lines were more resistant to P sojae, whereas other one had no significant difference in comparison with the control. These transgenic soybean lines with enhanced resistance to P sojae will be useful in soybean resistance breeding.展开更多
本课题组在前期研究中,从多头绒泡菌中分离到一个SR蛋白激酶基因psrpk,并将其编码蛋白命名为PSRPK(SR protein kinase ofPhysarum Polycephalum).为分离PSRPK相关蛋白基因以了解PSRPK的功能,构建了转化率为2×106转化子/3μg pGADT7...本课题组在前期研究中,从多头绒泡菌中分离到一个SR蛋白激酶基因psrpk,并将其编码蛋白命名为PSRPK(SR protein kinase ofPhysarum Polycephalum).为分离PSRPK相关蛋白基因以了解PSRPK的功能,构建了转化率为2×106转化子/3μg pGADT7-Rec、密度为5.35×108cells/mL、滴度为2.34×109cfu/mL的多头绒泡菌酵母双杂交AD库.以PSRPK为饵蛋白筛选该文库得到接合率为41.18%的杂交酵母,在SD/-Leu/-Trp/-Ade/-H is培养板上筛选获得1476个杂交克隆,其中,X-gal滤纸显色呈强蓝色的克隆有342个.对大于500 bp的67个克隆测序获得35个cDNA片段,其中编码Plasm in C、branched-chain am inoacid am inotransferase(BCAT)类似蛋白、MSF1类似蛋白、m ixed-linked glucanase precursor类似蛋白、FCY1p类似蛋白和40S ribosomal protein S2类似蛋白等7个cDNA片段在阳性杂交酵母克隆中出现多次,其余仅出现一次.在35个cDNA片段的编码序列中有15个具有同源蛋白,31个编码序列的丝氨酸含量大于或等于6%,符合激酶底物的组成特征.展开更多
Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale pr...Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic conditions.However,plants have evolved a number of survival strategies,among them is the induction of various stress-responsive genes such as the ribosomal protein large(RPL)gene.The RPL gene families encode critical proteins,which alleviate the effects of drought and salt stress in plants.In this study,comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt stresses.Results:Based on the genome-wide evaluation,26,8,and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum,G.raimondii,and G.arboreum,respectively.Furthermore,through bioinformatics analysis,key cis-regulatory elements related to RPL14B genes were discovered.The Myb binding sites(MBS),abscisic acid-responsive element(ABRE),CAAT-box,TATA box,TGACG-motif,and CGTCA-motif responsive to methyl jasmonate,as well as the TCA-motif responsive to salicylic acid,were identified.Expression analysis revealed a key gene,Gh_D01G0234(RPL14B),with significantly higher induction levels was further evaluated through a reverse genetic approach.The knockdown of Gh_D01G0234(RPL14B)significantly affected the performance of cotton seedlings under drought/salt stress conditions,as evidenced by a substantial reduction in various morphological and physiological traits.Moreover,the level of the antioxidant enzyme was significantly reduced in VIGS-plants,while oxidant enzyme levels increased significantly,as demonstrated by the higher malondialdehyde concentration level.Conclusion:The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes,which are key in oxidizing the various oxidants.The key pathways need to be investigated and even as we exploit these genes in the developing of more stress-resilient cotton germplasms.展开更多
基金supported by the National Natural Science Foundation of China(32321001)the Forestry Bureau of Anhui Province(AHLYJBGS-2024-01)+3 种基金the Center for Advanced Interdisciplinary Science and Biomedicine of IHM,the Division of Life Sciences and Medicine,the University of Science and Technology of China(QYPY20220012)the USTC Research Funds of the Double First-Class Initiative(YD9100002016)start-up funding from the University of Science and Technology of China and the Chinese Academy of Sciences(GG9100007007,KY9100000026,KY9100000051,KJ2070000079)the Fundamental Research Funds for the Central Universities(WK9100000021)。
文摘Protein biosynthesis by the ribosome is a fundamental biological process in living systems.Recent studies suggest that ribosomal subunits also play essential roles in cell growth and differentiation beyond their roles in protein translation.The ribosomal subunit RPS6 has been studied for more than 50 years in various organisms,but little is known about its specific roles in certain signaling pathways.In this study,we focused on the functions of Arabidopsis RPS6A in auxin-related root growth and development.The rps6a mutant presented a series of auxin-deficient phenotypes,such as shortened primary roots,reduced lateral root numbers,and defective vasculatures.Treatment of the rps6a mutant with various concentrations of auxin and its analogs did not restore the root defect phenotypes,suggesting a defect in the auxin signaling pathway.Further cell biological and global transcriptome analyses revealed that auxin signaling was weakened in the rps6a mutant and that there was a reduced abundance of PIN-FORMED(PIN)auxin transporters.Our work provides insights into the role of the protein biosynthesis pathway involved in auxin signaling.
文摘Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role.Methods Initially,we assessed PES1 expression patterns in two distinct senescence models:replicative senescent mouse embryonic fibroblasts(MEFs)and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells.Subsequently,PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types.Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays,respectively.The expression of senescence-associated proteins(p53,p21,and Rb)and SASP factors(IL-6,IL-1β,and IL-8)were analyzed by Western blot or qPCR.Furthermore,Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology.Results PES1 expression was significantly downregulated in senescent MEFs and HepG2 cells.PES1 knockdown resulted in decreased EdU-positive cells and increased SA-β-gal-positive cells,indicating proliferation inhibition and senescence induction.Mechanistically,PES1 suppression activated the p53-p21 pathway without affecting Rb expression,while upregulating IL-6,IL-1β,and IL-8 production.Notably,PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress,as evidenced by aberrant nucleolar morphology.Conclusion Our findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent(but Rb-independent)cellular senescence,highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.
文摘背景与目的:研究发现核糖体蛋白L8(ribosomal protein L8,RPL8)在黑色素瘤中表达能激活黑色素瘤患者外周血单个核细胞增殖并产生白细胞介素2,提示RPL8可能参与抗肿瘤免疫应答,有望成为抗肿瘤治疗的靶点。本研究通过RPL8蛋白负载树突状细胞(dentritic cell,DC),探讨负载RPL8蛋白的DC对黑色素瘤的免疫效应。方法:原核表达RPL8蛋白,纯化后致敏小鼠骨髓来源DC,流式细胞仪检测DC表面标志,MTT法检测细胞毒性T淋巴细胞对小鼠黑色素瘤细胞的杀伤作用;负载RPL8蛋白DC免疫治疗小鼠后,观察肿瘤体积变化及小鼠生存时间。结果:纯化蛋白经蛋白质印迹法(Western blot)分析见约28×103大小的特异性条带;DC经RPL8及细菌脂多糖(Lipoplysaccharide,LPS)诱导成熟后细胞表面CD11c、CD80、MHC-Ⅰ类、MHC-Ⅱ类分子表达增高,能激活T淋巴细胞,对B16细胞有抑制作用,RPL8-DC组的抑制率在效靶比为30∶1时高达70%,较PBS组和DC组明显增高;负载RPL8蛋白DC免疫治疗小鼠后,肿瘤体积缩小,小鼠的生存期明显延长。结论:负载RPL8的DC对黑色素瘤有生长抑制作用。
文摘Objective: In order to coordinately express amarker gene and therapeutic genes in one vector, apolycistronic retroviral vector was constructed withthe internal ribosome entry site from encephalo-myocarditis and polio virus, which was able to expressthree different genes simultaneously. Methods: Thevector used mouse moloney leukemia virus sequencesfor viral expression and packaging functions
文摘Interleukin-12, a heterodimer produced mainlyby antigen presenting cells (APCs), plays vital rolesin the induction of Thl-mediated immune responsesand has the potential to treat cancer and infectiousdiseases. Systemic administration of IL-12 wasobserved to be associated with severe toxicity. To takefull advantage of its immunoregulatory activities andminimize its toxicities, local administration of IL-12may be the ideal approach. Intratumoral injection
基金Supported by the National Items of Research and Industrial Development of Transgenic Plants(J99-B-013)
文摘Soybean is one of the most important sources of edible oil and proteins in the world. However, it suffers from many kinds of fungal diseases which is a major limiting factor in soybean production. The fungal disease can be effectively controlled by breeding plant cultivars with genetic transformation. In this study, the resistance to Phytophthora sojae of five bivalent transgenic soybean lines was identified using the hypocotyls inoculation technique. The lines were the T2 of the transgenic soybean which were transformed with kidney bean chitinase gene and barley ribosome inactivating protein gene, and were positive by Southern Blot analysis. The resistance difference was studied through comparing the death percentage of transgenic soybean with the control. The results showed that four lines were more resistant to P sojae, whereas other one had no significant difference in comparison with the control. These transgenic soybean lines with enhanced resistance to P sojae will be useful in soybean resistance breeding.
文摘本课题组在前期研究中,从多头绒泡菌中分离到一个SR蛋白激酶基因psrpk,并将其编码蛋白命名为PSRPK(SR protein kinase ofPhysarum Polycephalum).为分离PSRPK相关蛋白基因以了解PSRPK的功能,构建了转化率为2×106转化子/3μg pGADT7-Rec、密度为5.35×108cells/mL、滴度为2.34×109cfu/mL的多头绒泡菌酵母双杂交AD库.以PSRPK为饵蛋白筛选该文库得到接合率为41.18%的杂交酵母,在SD/-Leu/-Trp/-Ade/-H is培养板上筛选获得1476个杂交克隆,其中,X-gal滤纸显色呈强蓝色的克隆有342个.对大于500 bp的67个克隆测序获得35个cDNA片段,其中编码Plasm in C、branched-chain am inoacid am inotransferase(BCAT)类似蛋白、MSF1类似蛋白、m ixed-linked glucanase precursor类似蛋白、FCY1p类似蛋白和40S ribosomal protein S2类似蛋白等7个cDNA片段在阳性杂交酵母克隆中出现多次,其余仅出现一次.在35个cDNA片段的编码序列中有15个具有同源蛋白,31个编码序列的丝氨酸含量大于或等于6%,符合激酶底物的组成特征.
基金The National Natural Science Foundation of China(31621005,31530053,and 31671745)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences financially sponsored this research program.
文摘Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic conditions.However,plants have evolved a number of survival strategies,among them is the induction of various stress-responsive genes such as the ribosomal protein large(RPL)gene.The RPL gene families encode critical proteins,which alleviate the effects of drought and salt stress in plants.In this study,comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt stresses.Results:Based on the genome-wide evaluation,26,8,and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum,G.raimondii,and G.arboreum,respectively.Furthermore,through bioinformatics analysis,key cis-regulatory elements related to RPL14B genes were discovered.The Myb binding sites(MBS),abscisic acid-responsive element(ABRE),CAAT-box,TATA box,TGACG-motif,and CGTCA-motif responsive to methyl jasmonate,as well as the TCA-motif responsive to salicylic acid,were identified.Expression analysis revealed a key gene,Gh_D01G0234(RPL14B),with significantly higher induction levels was further evaluated through a reverse genetic approach.The knockdown of Gh_D01G0234(RPL14B)significantly affected the performance of cotton seedlings under drought/salt stress conditions,as evidenced by a substantial reduction in various morphological and physiological traits.Moreover,the level of the antioxidant enzyme was significantly reduced in VIGS-plants,while oxidant enzyme levels increased significantly,as demonstrated by the higher malondialdehyde concentration level.Conclusion:The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes,which are key in oxidizing the various oxidants.The key pathways need to be investigated and even as we exploit these genes in the developing of more stress-resilient cotton germplasms.
