Background:Breakpoint Cluster Region-Abelson(BCR::ABL1)fusion protein is essential in the pathogenesis of chronic myeloid leukemia(CML);however,the chronic-to-blast phase transformation remains elusive.We identified n...Background:Breakpoint Cluster Region-Abelson(BCR::ABL1)fusion protein is essential in the pathogenesis of chronic myeloid leukemia(CML);however,the chronic-to-blast phase transformation remains elusive.We identified novel kinesin light chain 2(KLC2)mutations in CML-myeloid blast phase patients.We aimed to examine the functional role of KLC2 mutations in leukemogenesis.Methods:To evaluate the biological role of KLC2 mutants(MT)in CML cells,we expressed KLC2-MT in different human CML cell lines harboring BCR::ABL1 and performed immunoblot,immunofluorescence,cell proliferation,differentiation,and apoptosis;Tyrosine kinase inhibitor(TKI)-drug activities;and clonogenic assays for in vitro functional analyses.We co-expressed KLC2-MT and BCR::ABL1 in mouse bone marrow cells(BMCs)to evaluate their clonogenic and self-renewal abilities ex vivo.Furthermore,we examined tumorigenic activity and drug efficacy in the K562 xenograft model.Results:KLC2-MT overexpression in BCR::ABL1-positive K562 and KU812 CML cells promoted cell proliferation and clonogenic potential,decreased imatinib sensitivity,and reduced apoptosis.Serial colony replating assays revealed that KLC2-MT and BCR::ABL1 co-expression enhanced the self-renewal ability of mouse BMCs with immature morphology.In the K562 xenograft model,KLC2-MT enhanced tumorigenic potential and diminished imatinib efficacy.Further studies reported that KLC2-MT augmented signal transducer and activator of transcription 3(STAT3)activation and nuclear accumulation in imatinib-treated CML cells.KLC2-WT and KLC2-MT interacted with mothers against decapentaplegic homolog 2(SMAD2);however,the latter impaired transforming growth factor-beta(TGF-β)–mediated SMAD2/3 activation while enhancing STAT3 phosphorylation.Conclusions:This study demonstrates the biological and functional importance of KLC2 mutation in CML cells,potentially enabling the development of better treatment strategies for CML patients carrying KLC2 mutations and providing enhanced understanding of the disease progression.展开更多
Calcium(Ca^(2+)), a crucial second messenger in plants, is involved in diverse signaling pathways including biotic and abiotic stress responses. However, the functions of most calcium sensors including calcium-depende...Calcium(Ca^(2+)), a crucial second messenger in plants, is involved in diverse signaling pathways including biotic and abiotic stress responses. However, the functions of most calcium sensors including calcium-dependent protein kinases(CDPKs), calmodulins(Ca Ms), calmodulinlike proteins(CMLs), and calcineurin B-like proteins(CBLs) remain limited in plants, particularly in woody plants. Currently, a total of 83 CMLs and seven Ca Ms were discovered in the apple genome in this study. Functional domain analysis showed that these CMLs and Ca Ms contain a highly conserved EF-hand motif. q RT-PCR assays indicated that these CMLs were expressed ubiquitously in apple, including in the roots, stems,leaves, flowers, and fruits, and also possessed spatial specificity. Finally, most of these CMLs were induced by hormonal and abiotic stress, suggesting their potential roles in the regulation of growth, development, and the stress response in apple. In summary, our findings provide potential evidence that apple CMLs may be involved in abiotic stress and the regulation of plant growth and development.展开更多
Calmodulin-like (CMLs) proteins are critical in calcium signaling and essential for plant growth, development, and stress responses. In many species, the CMLs families have been identified and described. However, the ...Calmodulin-like (CMLs) proteins are critical in calcium signaling and essential for plant growth, development, and stress responses. In many species, the CMLs families have been identified and described. However, the characterization and expression profiling of CMLs genes in tobacco is retrievable. In this study, a comprehensive whole-genome identification and analysis, and 75 NtCML genes were identified in tobacco, each containing two to four EF-hand domains. Most NtCML proteins exhibited conserved gene structures and motifs. Notably, most NtCML proteins were intron-less and distributed across 18 chromosomes. Two pairs of tandemly duplicated genes and seven pairs of segmentally duplicated genes were identified within the tobacco genome. Furthermore, 22 pairs of orthologous CMLs genes were discovered between Arabidopsis and tobacco. Cis-acting element analysis revealed that elements associated with hormones, stress responses, and plant growth and development were found in the promoter regions. Expression analysis indicated that some NtCML genes displayed tissue-specific expression patterns. Specifically, NtCML12, NtCML18, NtCML27, and NtCML28 showed significant upregulation during cold acclimation treatment. These results indicate that tobacco CMLs act as Ca^(2+) signal transducers, regulating plant growth and abiotic stress responses.展开更多
【目的】鉴定青海茄参(Mandragora chinghaiensis Kuang et A.M.Jl)CML基因家族成员,探究其对冷胁迫的响应。【方法】基于课题组的全长转录组数据,利用生物信息学技术对青海茄参CML基因家族的成员及其蛋白理化性质、系统发育过程、基因...【目的】鉴定青海茄参(Mandragora chinghaiensis Kuang et A.M.Jl)CML基因家族成员,探究其对冷胁迫的响应。【方法】基于课题组的全长转录组数据,利用生物信息学技术对青海茄参CML基因家族的成员及其蛋白理化性质、系统发育过程、基因结构等进行分析。【结果】在青海茄参中共鉴定出19个CML基因家族成员,亚细胞定位主要在细胞核,含有1~4个EF-hand结构域,二三级结构主要由α-螺旋组成;基序分析发现,McCMLs蛋白含有4个典型的EF手部基序;系统进化树分析显示,McCMLs蛋白被分为3个亚族。冷胁迫表达模式分析表明,McCML基因家族在冷胁迫下差异表达,其中McCML3、McCML7表达量显著上调。【结论】通过对McCML基因家族进行鉴定和分析,筛选出McCML3、McCML7作为青海茄参抗寒候选基因,为后期研究其抗寒分子机制提供理论基础。展开更多
基金supported by grants from the Ministry of Science and Technology,Taiwan(MOST108-2314-B-182-006,MOST109-2314-B-182-071:Lee-Yung Shih)the Ministry of Health and Welfare,Taiwan(MOHW110-TDU-B-212-134011:Lee-Yung Shih)+3 种基金Chang Gung Memorial Hospital(CMRPG3D1524,OMRPG3E0031:Lee-Yung Shih)the Grant-in-Aid for the Japan Society for the Promotion of Science(JSPS)KAKENHI(JP19H05656:Seishi Ogawa,22K16320:Yotaro Ochi)the Japan Agency for Medical Research and Development(AMED)(JP19cm0106501h0004,JP19ck0106250h0003:Seishi Ogawa)the Ministry of Education,Culture,Sports,Science and Technology of Japan(MEXT)(hp200138,hp210167:Seishi Ogawa)。
文摘Background:Breakpoint Cluster Region-Abelson(BCR::ABL1)fusion protein is essential in the pathogenesis of chronic myeloid leukemia(CML);however,the chronic-to-blast phase transformation remains elusive.We identified novel kinesin light chain 2(KLC2)mutations in CML-myeloid blast phase patients.We aimed to examine the functional role of KLC2 mutations in leukemogenesis.Methods:To evaluate the biological role of KLC2 mutants(MT)in CML cells,we expressed KLC2-MT in different human CML cell lines harboring BCR::ABL1 and performed immunoblot,immunofluorescence,cell proliferation,differentiation,and apoptosis;Tyrosine kinase inhibitor(TKI)-drug activities;and clonogenic assays for in vitro functional analyses.We co-expressed KLC2-MT and BCR::ABL1 in mouse bone marrow cells(BMCs)to evaluate their clonogenic and self-renewal abilities ex vivo.Furthermore,we examined tumorigenic activity and drug efficacy in the K562 xenograft model.Results:KLC2-MT overexpression in BCR::ABL1-positive K562 and KU812 CML cells promoted cell proliferation and clonogenic potential,decreased imatinib sensitivity,and reduced apoptosis.Serial colony replating assays revealed that KLC2-MT and BCR::ABL1 co-expression enhanced the self-renewal ability of mouse BMCs with immature morphology.In the K562 xenograft model,KLC2-MT enhanced tumorigenic potential and diminished imatinib efficacy.Further studies reported that KLC2-MT augmented signal transducer and activator of transcription 3(STAT3)activation and nuclear accumulation in imatinib-treated CML cells.KLC2-WT and KLC2-MT interacted with mothers against decapentaplegic homolog 2(SMAD2);however,the latter impaired transforming growth factor-beta(TGF-β)–mediated SMAD2/3 activation while enhancing STAT3 phosphorylation.Conclusions:This study demonstrates the biological and functional importance of KLC2 mutation in CML cells,potentially enabling the development of better treatment strategies for CML patients carrying KLC2 mutations and providing enhanced understanding of the disease progression.
基金grants from NSFC(31601728,31272142)Shandong Province(ZR2016CQ13,SDAIT-06-03)+1 种基金Young Scientists Funds of Shandong Agricultural University(564024)Youth Science and Technology Innovation Fund of Shandong Agricultural University(24024)
文摘Calcium(Ca^(2+)), a crucial second messenger in plants, is involved in diverse signaling pathways including biotic and abiotic stress responses. However, the functions of most calcium sensors including calcium-dependent protein kinases(CDPKs), calmodulins(Ca Ms), calmodulinlike proteins(CMLs), and calcineurin B-like proteins(CBLs) remain limited in plants, particularly in woody plants. Currently, a total of 83 CMLs and seven Ca Ms were discovered in the apple genome in this study. Functional domain analysis showed that these CMLs and Ca Ms contain a highly conserved EF-hand motif. q RT-PCR assays indicated that these CMLs were expressed ubiquitously in apple, including in the roots, stems,leaves, flowers, and fruits, and also possessed spatial specificity. Finally, most of these CMLs were induced by hormonal and abiotic stress, suggesting their potential roles in the regulation of growth, development, and the stress response in apple. In summary, our findings provide potential evidence that apple CMLs may be involved in abiotic stress and the regulation of plant growth and development.
基金College of Agronomy and Biotechnology,Southwest University(4412200577).
文摘Calmodulin-like (CMLs) proteins are critical in calcium signaling and essential for plant growth, development, and stress responses. In many species, the CMLs families have been identified and described. However, the characterization and expression profiling of CMLs genes in tobacco is retrievable. In this study, a comprehensive whole-genome identification and analysis, and 75 NtCML genes were identified in tobacco, each containing two to four EF-hand domains. Most NtCML proteins exhibited conserved gene structures and motifs. Notably, most NtCML proteins were intron-less and distributed across 18 chromosomes. Two pairs of tandemly duplicated genes and seven pairs of segmentally duplicated genes were identified within the tobacco genome. Furthermore, 22 pairs of orthologous CMLs genes were discovered between Arabidopsis and tobacco. Cis-acting element analysis revealed that elements associated with hormones, stress responses, and plant growth and development were found in the promoter regions. Expression analysis indicated that some NtCML genes displayed tissue-specific expression patterns. Specifically, NtCML12, NtCML18, NtCML27, and NtCML28 showed significant upregulation during cold acclimation treatment. These results indicate that tobacco CMLs act as Ca^(2+) signal transducers, regulating plant growth and abiotic stress responses.
文摘【目的】鉴定青海茄参(Mandragora chinghaiensis Kuang et A.M.Jl)CML基因家族成员,探究其对冷胁迫的响应。【方法】基于课题组的全长转录组数据,利用生物信息学技术对青海茄参CML基因家族的成员及其蛋白理化性质、系统发育过程、基因结构等进行分析。【结果】在青海茄参中共鉴定出19个CML基因家族成员,亚细胞定位主要在细胞核,含有1~4个EF-hand结构域,二三级结构主要由α-螺旋组成;基序分析发现,McCMLs蛋白含有4个典型的EF手部基序;系统进化树分析显示,McCMLs蛋白被分为3个亚族。冷胁迫表达模式分析表明,McCML基因家族在冷胁迫下差异表达,其中McCML3、McCML7表达量显著上调。【结论】通过对McCML基因家族进行鉴定和分析,筛选出McCML3、McCML7作为青海茄参抗寒候选基因,为后期研究其抗寒分子机制提供理论基础。
