Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeepin...Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeeping functions,they are also known to be involved in immune response during bacterial infection.In this study,we identified and annotated 10 hsp10/60 genes through bioinformatic analysis in Japanese flounder(Paralichthys olivaceus).Among them one member of hsp10(hspe)family and nine members of hsp60(hspd)family were identified.Phylogenetic and selection pressure analysis showed that the hsp10/60 genes were evolutionarily constrained and their function was conserved.Besides,hsp10/60 genes were involved in different embryonic and larval stages and acted as the sentinel role in an unchallenged organism.In addition,we also observed the expression patterns of hsp10/60 genes after Edwardsiella tarda infection,for the first time in Japanese flounder.Eight out of 10 genes were differentially expressed after bacterial challenges,the significantly regulated expressions of flounder hsp10/60 genes after bacterial infections suggested their involvement in immune response in flounder.Our results provide valuable information for clarifying the evolutionary relationship,and early insights of the immune functions of hsp10/60 genes in Japanese flounder.展开更多
背景与目的：研究热休克蛋白60基因（heat shock protein 60 gene,Hsp60）在小鼠胚胎腭、前肢正常和异常发育过程中的表达情况。材料与方法：将受孕后ICR小鼠随机分为实验组和对照组2组,每组各64只,于孕10d（gestational day 10,GD10）...背景与目的：研究热休克蛋白60基因（heat shock protein 60 gene,Hsp60）在小鼠胚胎腭、前肢正常和异常发育过程中的表达情况。材料与方法：将受孕后ICR小鼠随机分为实验组和对照组2组,每组各64只,于孕10d（gestational day 10,GD10）,分别经口一次给予实验组孕鼠80mg/kg的全反式视黄酸,对照组孕鼠给予等体积的大豆油,并分别于GD11-GD18取2组胎鼠的前肢,于GD15-GD17取2组胎鼠的腭,利用实时荧光定量PCR检测Hsp60的表达丰度。结果：Hsp60在正常、异常肢和腭中均有表达。对照肢的表达丰度在GD14和出生前呈高表达,而实验肢的表达在各胚龄无明显差异（P〉0.05）;实验肢Hsp60在GD11-GD18的表达水平高于同一胚龄的对照肢（P〈0.05）。在正常腭中,Hsp60恒定表达,在异常腭中,Hsp60的表达随胚龄增大而降低;在GD15-GD17的表达丰度为实验腭低于同胚龄的对照腭（P〈0.05）。结论：全反式视黄酸所致的短肢中,Hsp60的表达呈应激性升高;而在异常腭中,Hsp60的表达受到抑制。展开更多
本研究采用RT-PCR技术克隆了药材甲热激蛋白60(Heat shock protein 60,Hsp60)基因的cDNA全长序列,命名为SpHsp60(GenBank登录号:KX778623).氨基酸序列分析表明SpHsp60具有Hsp60基因家族高度保守的基序,实时定量PCR技术检测了不同温度胁...本研究采用RT-PCR技术克隆了药材甲热激蛋白60(Heat shock protein 60,Hsp60)基因的cDNA全长序列,命名为SpHsp60(GenBank登录号:KX778623).氨基酸序列分析表明SpHsp60具有Hsp60基因家族高度保守的基序,实时定量PCR技术检测了不同温度胁迫后该基因的表达量,结果表明:-5℃和0℃低温处理2h,药材甲成虫体内的SpHsp60的表达量均显著高于对照组,且42℃高温处理2h后的表达量也显著高于对照组.说明SpHSP60与药材甲应对极端温度胁迫相关.展开更多
The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a ch...The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.展开更多
基金This work was supported by the National Key Research and Development Program of China(No.2018YFD0900601)the Natural Science Foundation of Shandong Province(No.ZR2017MC072).
文摘Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeeping functions,they are also known to be involved in immune response during bacterial infection.In this study,we identified and annotated 10 hsp10/60 genes through bioinformatic analysis in Japanese flounder(Paralichthys olivaceus).Among them one member of hsp10(hspe)family and nine members of hsp60(hspd)family were identified.Phylogenetic and selection pressure analysis showed that the hsp10/60 genes were evolutionarily constrained and their function was conserved.Besides,hsp10/60 genes were involved in different embryonic and larval stages and acted as the sentinel role in an unchallenged organism.In addition,we also observed the expression patterns of hsp10/60 genes after Edwardsiella tarda infection,for the first time in Japanese flounder.Eight out of 10 genes were differentially expressed after bacterial challenges,the significantly regulated expressions of flounder hsp10/60 genes after bacterial infections suggested their involvement in immune response in flounder.Our results provide valuable information for clarifying the evolutionary relationship,and early insights of the immune functions of hsp10/60 genes in Japanese flounder.
文摘背景与目的：研究热休克蛋白60基因（heat shock protein 60 gene,Hsp60）在小鼠胚胎腭、前肢正常和异常发育过程中的表达情况。材料与方法：将受孕后ICR小鼠随机分为实验组和对照组2组,每组各64只,于孕10d（gestational day 10,GD10）,分别经口一次给予实验组孕鼠80mg/kg的全反式视黄酸,对照组孕鼠给予等体积的大豆油,并分别于GD11-GD18取2组胎鼠的前肢,于GD15-GD17取2组胎鼠的腭,利用实时荧光定量PCR检测Hsp60的表达丰度。结果：Hsp60在正常、异常肢和腭中均有表达。对照肢的表达丰度在GD14和出生前呈高表达,而实验肢的表达在各胚龄无明显差异（P〉0.05）;实验肢Hsp60在GD11-GD18的表达水平高于同一胚龄的对照肢（P〈0.05）。在正常腭中,Hsp60恒定表达,在异常腭中,Hsp60的表达随胚龄增大而降低;在GD15-GD17的表达丰度为实验腭低于同胚龄的对照腭（P〈0.05）。结论：全反式视黄酸所致的短肢中,Hsp60的表达呈应激性升高;而在异常腭中,Hsp60的表达受到抑制。
文摘本研究采用RT-PCR技术克隆了药材甲热激蛋白60(Heat shock protein 60,Hsp60)基因的cDNA全长序列,命名为SpHsp60(GenBank登录号:KX778623).氨基酸序列分析表明SpHsp60具有Hsp60基因家族高度保守的基序,实时定量PCR技术检测了不同温度胁迫后该基因的表达量,结果表明:-5℃和0℃低温处理2h,药材甲成虫体内的SpHsp60的表达量均显著高于对照组,且42℃高温处理2h后的表达量也显著高于对照组.说明SpHSP60与药材甲应对极端温度胁迫相关.
文摘The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.
