The mitogen-activated protein kinase(MAPK)cascade is crucial to plant growth,development,and stress responses.MAPK kinases(MAPKK)play a vital role in linking upstream MAPKK kinases(MAPKKK)with the downstream MAPK.Blac...The mitogen-activated protein kinase(MAPK)cascade is crucial to plant growth,development,and stress responses.MAPK kinases(MAPKK)play a vital role in linking upstream MAPKK kinases(MAPKKK)with the downstream MAPK.Black spot is one of the most serious fungal diseases of pear which is an important part of the fruit industry in China.The MAPKK genes have been identified in many plants,however,none has been reported in pear(Pyrus bretschneideri).In order to explore whether MAPK gene of pear is related to black spot disease,we designed this experiment.The present study investigated eight putative PbrMAPKK genes obtained from the Chinese white pear genome.The phylogenetic analysis revealed that PbrMAPKK genes were divided into A,B,C,and D groups.These PbrMAPKK genes are randomly distributed on 7 out of 17 chromosomes and mainly originated from the whole-genome duplication(WGD)event.The expression analysis of PbrMAPKK genes in seven pear tissues and the leaves of susceptible and resistant varieties after Alternaria alternata infection by quantitative real-time PCR(qRT-PCR)identified seven candidate genes associated with resistance.Furthermore,virus-induced gene silencing(VIGS)indicated that PbrMAPKK6 gene enhanced resistance to pear black spot disease in pear.展开更多
Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein ki...Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein kinase known as the CBL-interacting protein kinase(CIPK).In plants,the CBL-CIPK signaling network regulates multiple signals in response to different extracellular cues including abiotic stress.However,the genome wide annotation and expression patterns of CBLs and CIPKs in woody cutting flower plants are still unclear.In this study,a total number of 7 CBLs(RcCBLs)and 17 CIPKs(RcCIPKs)genes,divided into four and five subfamilies,respectively,were identified from the rose genome.All RcCBLs possess a classic elongation factor-hand(EF-hand)domain,while all RcCIPKs possess both the classic kinase and NAF domains.Most RcCBLs were predicted to be plasma membrane localized,whereas most RcCIPKs were predicted to be cytoplasmic localized.Synteny analysis showed that one RcCBL gene pair and five RcCIPK gene pairs have gone through whole genome duplication events.Promoter cis-element prediction assays indicated that RcCBLs and RcCIPKs could function in different abiotic stress responses in rose plants.Further quantitative real-time PCR analysis demonstrated that RcCBLs and RcCIPKs were expressed in different organs with overlapped but distinct patterns in response to various abiotic stresses.The findings in this work will provide fundamental information and gene resources for further functional research on RcCBLs and RcCIPKs.展开更多
Background:Microribose nucleic acids(miRNAs)are implicated in the progression of lung adenocarcinoma.MicroRNA-345-5p(miR-345-5p)is a recently identified anti-oncogene in some human cancers,but its functional role and ...Background:Microribose nucleic acids(miRNAs)are implicated in the progression of lung adenocarcinoma.MicroRNA-345-5p(miR-345-5p)is a recently identified anti-oncogene in some human cancers,but its functional role and possible molecular mechanism in lung adenocarcinoma remain unknown.This study aimed to identify the biological function and underlying mechanism of miR-345-5p in lung adenocarcinoma cells.Methods:In this study,lung adenocarcinoma tissues and adjacent tissues were collected in the First Affiliated Hospital of Anhui Medical University between April 2016 and February 2017.The expression of miR-345-5p and ras homolog family member A(RhoA)in lung adenocarcinoma tissues and human lung adenocarcinoma cell lines(A549,H1650,PC-9,and H441)was detected by reverse transcription quantitative polymerase chain reaction analysis.Functional assays including colony formation,flow cytometry analysis,wound healing,and transwell assays were performed to assess the proliferation,apoptosis,migration,and invasion of lung adenocarcinoma cells.In addition,RNA pulldown and luciferase reporter assays were conducted to evaluate the relationship between miR-345-5p and RhoA.Difference between the two groups was analyzed with Student’st test,while that among multiple groups was analyzed with one-way analysis of variance.Results:MiR-345-5p expression displayed lower level in lung adenocarcinoma tissues(0.241±0.095vs.1.000±0.233,t=19.247,P<0.001)and cell lines(F=56.992,P<0.001)than control tissues and cells.Functional experiments demonstrated that upregulation of miR-345-5p inhibited the malignant phenotypes of lung adenocarcinoma cells via suppressing cell proliferation,migration,invasion,and facilitating cell apoptosis.Additionally,RhoA was verified to be the downstream target of miR-345-5p.Expression of RhoA was downregulated by overexpression of miR-345-5p in PC-9(0.321±0.047vs.1.000±0.127,t=8.536,P<0.001)and H1650(0.398±0.054vs.1.000±0.156,t=4.429,P=0.011)cells.Rescue assays revealed that overexpression of RhoA rescued the suppressive effects of miR-345-5p upregulation on proliferation,migration,and invasion of lung adenocarcinoma cells.Further,miR-345-5p was found to regulate the Rho/Rho-associated protein kinase(ROCK)signaling pathway by downregulation of RhoA in lung adenocarcinoma cells.Conclusions:MiR-345-5p plays a tumor suppressor role in lung adenocarcinoma cells by downregulating RhoA to inactivate the Rho/ROCK pathway.展开更多
为了阐明大菱鲆丝裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinases,MAPKKs或MKKs)基因家族在生物和非生物应激响应中的作用,本实验首先通过生物信息学方法对大菱鲆MKK基因家族进行了全基因组水平的鉴定,利用多个应激相...为了阐明大菱鲆丝裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinases,MAPKKs或MKKs)基因家族在生物和非生物应激响应中的作用,本实验首先通过生物信息学方法对大菱鲆MKK基因家族进行了全基因组水平的鉴定,利用多个应激相关转录组数据集分析了大菱鲆MKK家族成员在不同组织及不同生物和非生物应激下的表达模式。结果显示,本研究在大菱鲆全基因组水平上共鉴定出9个MKK基因家族成员,它们不均匀地分布在7条染色体上,并分别对其编码蛋白的理化性质、蛋白二级结构和亚细胞定位进行了预测。基于系统发育分析,将SmMKKs划分为5个亚家族。内含-外显子结构、保守基序和多重序列比对分析结果不仅为大菱鲆MKK亚家族分类提供了证据,而且表明SmMKKs在进化上高度保守。SmMKKs在不同组织及不同生物和非生物应激下的基因表达模式分析表明,SmMKKs具有明显的组织特异性表达。另外,结果显示,粘孢子虫和肿大细胞病毒感染后,SmMKK6a呈极显著差异表达;热应激处理后,SmMKK6a呈极显著差异表达;高盐或低盐胁迫后,SmMKK4a、SmMKK4b、SmMKK6a和SmMKK7呈极显著差异表达。SmMKK6a在各种应激条件下均表现出极显著响应,表明其可能在综合抗应激中具有潜在的作用。这可能是第一个对大菱鲆MKK基因家族进行系统识别和功能分析的研究。以上研究结果不仅表明MKK基因家族在大菱鲆响应多种生物和非生物应激中发挥重要作用,而且也为大菱鲆综合抗逆分子选择育种研究提供了重要的理论支撑。展开更多
Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibi...Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibitory environment for axonal regeneration. Among these inhibitory molecules, myelinassociated inhibitors, including neurite outgrowth inhibitor A, oligodendrocyte myelin glycoprotein, myelin-associated glycoprotein, chondroitin sulfate proteoglycans and repulsive guidance molecule A are of particular importance. Due to their inhibitory nature, they represent exciting molecular targets to study axonal inhibition and regeneration after central injuries. These molecules are mainly produced by neurons, oligodendrocytes, and astrocytes within the scar and in its immediate vicinity. They exert their effects by binding to specific receptors, localized in the membranes of neurons. Receptors for these inhibitory cues include Nogo receptor 1, leucine-rich repeat, and Ig domain containing 1 and p75 neurotrophin receptor/tumor necrosis factor receptor superfamily member 19(that form a receptor complex that binds all myelin-associated inhibitors), and also paired immunoglobulin-like receptor B. Chondroitin sulfate proteoglycans and repulsive guidance molecule A bind to Nogo receptor 1, Nogo receptor 3, receptor protein tyrosine phosphatase σ and leucocyte common antigen related phosphatase, and neogenin, respectively. Once activated, these receptors initiate downstream signaling pathways, the most common amongst them being the Rho A/ROCK signaling pathway. These signaling cascades result in actin depolymerization, neurite outgrowth inhibition, and failure to regenerate after spinal cord injury. Currently, there are no approved pharmacological treatments to overcome spinal cord injuries other than physical rehabilitation and management of the array of symptoms brought on by spinal cord injuries. However, several novel therapies aiming to modulate these inhibitory proteins and/or their receptors are under investigation in ongoing clinical trials. Investigation has also been demonstrating that combinatorial therapies of growth inhibitors with other therapies, such as growth factors or stem-cell therapies, produce stronger results and their potential application in the clinics opens new venues in spinal cord injury treatment.展开更多
基金supported by the National Key Research and Development Program of China(Grant No.2022YFD1200503)Jiangsu Agriculture Science and Technology Innovation Fund[Grant Nos.SCX(22)3215],Fundamental Research Funds for the Central Universities(Grant No.JCQY201901)the Earmarked Fund for China Agriculture Research System(Grant No.CARS-28).
文摘The mitogen-activated protein kinase(MAPK)cascade is crucial to plant growth,development,and stress responses.MAPK kinases(MAPKK)play a vital role in linking upstream MAPKK kinases(MAPKKK)with the downstream MAPK.Black spot is one of the most serious fungal diseases of pear which is an important part of the fruit industry in China.The MAPKK genes have been identified in many plants,however,none has been reported in pear(Pyrus bretschneideri).In order to explore whether MAPK gene of pear is related to black spot disease,we designed this experiment.The present study investigated eight putative PbrMAPKK genes obtained from the Chinese white pear genome.The phylogenetic analysis revealed that PbrMAPKK genes were divided into A,B,C,and D groups.These PbrMAPKK genes are randomly distributed on 7 out of 17 chromosomes and mainly originated from the whole-genome duplication(WGD)event.The expression analysis of PbrMAPKK genes in seven pear tissues and the leaves of susceptible and resistant varieties after Alternaria alternata infection by quantitative real-time PCR(qRT-PCR)identified seven candidate genes associated with resistance.Furthermore,virus-induced gene silencing(VIGS)indicated that PbrMAPKK6 gene enhanced resistance to pear black spot disease in pear.
基金funded by the following grants:the Natural Science Foundation of Shandong Province of China(ZR2021MC169)the Cooperation Project of University and Local Enterprise in Yantai of Shandong Province(2021XDRHXMPT09).
文摘Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein kinase known as the CBL-interacting protein kinase(CIPK).In plants,the CBL-CIPK signaling network regulates multiple signals in response to different extracellular cues including abiotic stress.However,the genome wide annotation and expression patterns of CBLs and CIPKs in woody cutting flower plants are still unclear.In this study,a total number of 7 CBLs(RcCBLs)and 17 CIPKs(RcCIPKs)genes,divided into four and five subfamilies,respectively,were identified from the rose genome.All RcCBLs possess a classic elongation factor-hand(EF-hand)domain,while all RcCIPKs possess both the classic kinase and NAF domains.Most RcCBLs were predicted to be plasma membrane localized,whereas most RcCIPKs were predicted to be cytoplasmic localized.Synteny analysis showed that one RcCBL gene pair and five RcCIPK gene pairs have gone through whole genome duplication events.Promoter cis-element prediction assays indicated that RcCBLs and RcCIPKs could function in different abiotic stress responses in rose plants.Further quantitative real-time PCR analysis demonstrated that RcCBLs and RcCIPKs were expressed in different organs with overlapped but distinct patterns in response to various abiotic stresses.The findings in this work will provide fundamental information and gene resources for further functional research on RcCBLs and RcCIPKs.
文摘Background:Microribose nucleic acids(miRNAs)are implicated in the progression of lung adenocarcinoma.MicroRNA-345-5p(miR-345-5p)is a recently identified anti-oncogene in some human cancers,but its functional role and possible molecular mechanism in lung adenocarcinoma remain unknown.This study aimed to identify the biological function and underlying mechanism of miR-345-5p in lung adenocarcinoma cells.Methods:In this study,lung adenocarcinoma tissues and adjacent tissues were collected in the First Affiliated Hospital of Anhui Medical University between April 2016 and February 2017.The expression of miR-345-5p and ras homolog family member A(RhoA)in lung adenocarcinoma tissues and human lung adenocarcinoma cell lines(A549,H1650,PC-9,and H441)was detected by reverse transcription quantitative polymerase chain reaction analysis.Functional assays including colony formation,flow cytometry analysis,wound healing,and transwell assays were performed to assess the proliferation,apoptosis,migration,and invasion of lung adenocarcinoma cells.In addition,RNA pulldown and luciferase reporter assays were conducted to evaluate the relationship between miR-345-5p and RhoA.Difference between the two groups was analyzed with Student’st test,while that among multiple groups was analyzed with one-way analysis of variance.Results:MiR-345-5p expression displayed lower level in lung adenocarcinoma tissues(0.241±0.095vs.1.000±0.233,t=19.247,P<0.001)and cell lines(F=56.992,P<0.001)than control tissues and cells.Functional experiments demonstrated that upregulation of miR-345-5p inhibited the malignant phenotypes of lung adenocarcinoma cells via suppressing cell proliferation,migration,invasion,and facilitating cell apoptosis.Additionally,RhoA was verified to be the downstream target of miR-345-5p.Expression of RhoA was downregulated by overexpression of miR-345-5p in PC-9(0.321±0.047vs.1.000±0.127,t=8.536,P<0.001)and H1650(0.398±0.054vs.1.000±0.156,t=4.429,P=0.011)cells.Rescue assays revealed that overexpression of RhoA rescued the suppressive effects of miR-345-5p upregulation on proliferation,migration,and invasion of lung adenocarcinoma cells.Further,miR-345-5p was found to regulate the Rho/Rho-associated protein kinase(ROCK)signaling pathway by downregulation of RhoA in lung adenocarcinoma cells.Conclusions:MiR-345-5p plays a tumor suppressor role in lung adenocarcinoma cells by downregulating RhoA to inactivate the Rho/ROCK pathway.
文摘为了阐明大菱鲆丝裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinases,MAPKKs或MKKs)基因家族在生物和非生物应激响应中的作用,本实验首先通过生物信息学方法对大菱鲆MKK基因家族进行了全基因组水平的鉴定,利用多个应激相关转录组数据集分析了大菱鲆MKK家族成员在不同组织及不同生物和非生物应激下的表达模式。结果显示,本研究在大菱鲆全基因组水平上共鉴定出9个MKK基因家族成员,它们不均匀地分布在7条染色体上,并分别对其编码蛋白的理化性质、蛋白二级结构和亚细胞定位进行了预测。基于系统发育分析,将SmMKKs划分为5个亚家族。内含-外显子结构、保守基序和多重序列比对分析结果不仅为大菱鲆MKK亚家族分类提供了证据,而且表明SmMKKs在进化上高度保守。SmMKKs在不同组织及不同生物和非生物应激下的基因表达模式分析表明,SmMKKs具有明显的组织特异性表达。另外,结果显示,粘孢子虫和肿大细胞病毒感染后,SmMKK6a呈极显著差异表达;热应激处理后,SmMKK6a呈极显著差异表达;高盐或低盐胁迫后,SmMKK4a、SmMKK4b、SmMKK6a和SmMKK7呈极显著差异表达。SmMKK6a在各种应激条件下均表现出极显著响应,表明其可能在综合抗应激中具有潜在的作用。这可能是第一个对大菱鲆MKK基因家族进行系统识别和功能分析的研究。以上研究结果不仅表明MKK基因家族在大菱鲆响应多种生物和非生物应激中发挥重要作用,而且也为大菱鲆综合抗逆分子选择育种研究提供了重要的理论支撑。
基金a Ph D fellowship by FCT-Fundacao para a Ciência Tecnologia (SFRH/BD/135868/2018)(to SSC)。
文摘Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibitory environment for axonal regeneration. Among these inhibitory molecules, myelinassociated inhibitors, including neurite outgrowth inhibitor A, oligodendrocyte myelin glycoprotein, myelin-associated glycoprotein, chondroitin sulfate proteoglycans and repulsive guidance molecule A are of particular importance. Due to their inhibitory nature, they represent exciting molecular targets to study axonal inhibition and regeneration after central injuries. These molecules are mainly produced by neurons, oligodendrocytes, and astrocytes within the scar and in its immediate vicinity. They exert their effects by binding to specific receptors, localized in the membranes of neurons. Receptors for these inhibitory cues include Nogo receptor 1, leucine-rich repeat, and Ig domain containing 1 and p75 neurotrophin receptor/tumor necrosis factor receptor superfamily member 19(that form a receptor complex that binds all myelin-associated inhibitors), and also paired immunoglobulin-like receptor B. Chondroitin sulfate proteoglycans and repulsive guidance molecule A bind to Nogo receptor 1, Nogo receptor 3, receptor protein tyrosine phosphatase σ and leucocyte common antigen related phosphatase, and neogenin, respectively. Once activated, these receptors initiate downstream signaling pathways, the most common amongst them being the Rho A/ROCK signaling pathway. These signaling cascades result in actin depolymerization, neurite outgrowth inhibition, and failure to regenerate after spinal cord injury. Currently, there are no approved pharmacological treatments to overcome spinal cord injuries other than physical rehabilitation and management of the array of symptoms brought on by spinal cord injuries. However, several novel therapies aiming to modulate these inhibitory proteins and/or their receptors are under investigation in ongoing clinical trials. Investigation has also been demonstrating that combinatorial therapies of growth inhibitors with other therapies, such as growth factors or stem-cell therapies, produce stronger results and their potential application in the clinics opens new venues in spinal cord injury treatment.
