To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal R...To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable informat展开更多
The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied ph...The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied physiological and biochemical processes especially in the stress response of plants. In this study, a total of 23 RB‐GRPs distributed on 10 chromosomes were identified in maize（Zea mays L.）, and they were divided into four subgroups according to their conserved domain architecture. Five pairs of paralogs were identified,while none of them was located on the same chromosomal region, suggesting that segmental duplication is predominant in the duplication events of the RB‐GRPs in maize. Comparative analysis of RB‐GRPs in maize, Arabidopsis（Arabidopsis thaliana L.）, rice（Oryza sativa L.）, and wheat（Triticum aestivum）revealed that two exclusive subgroups were only identified in maize. Expression of eight ZmRB‐GRPs was significantly regulated by at least two kinds of stresses. In addition, cis‐elements predicted in the promoter regions of the ZmRB‐GRPs also indicated that these ZmRB‐GRPs would be involved in stress response of maize. The preliminary genome‐wide analysis of the RB‐GRPs in maize would provide useful information for further study on the function of the ZmRB‐GRPs.展开更多
BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previo...BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previous study has demonstrated that CRP2 can be detected in the embryonic rat inner ear but not in the adult rat inner ear. However, at present, the expression of LIM protein family H members in stem or precursor cells has not been described. OBJECTIVE： To determine the expression and sub-cellular localization of CRP2 in olfactory stem cells. DESIGN, TIME AND SETTING： An experiment with repeated measures was performed in the Laboratory of Otorhinolaryngology, Head and Neck Surgery, Xijing Hospital, the Fourth Military Medical University from February 2008 to April 2008. MATERIALS： Olfactory stem cells, and rabbit-anti-CRP2 polyclonal antibody were prepared and kept in our laboratory. METHODS： Reverse transcription polymerase chain reaction and Western blot analysis were used to detect expression of CRP2 in olfactory stem cells. Immunocytochemistry was also used to localize CRP2 in olfactory stem cells. MAIN OUTCOME MEASURES： The expression and sub-cellular localization of CRP2 in rat olfactory stem cells. RESULTS： CRP2 expression was found in olfactory stem cells, and CRP2 was distributed in both the nucleus and the cytoplasm. CONCLUSION： Confirmation of the expression and distribution of CRP2 in olfactory stem cells.展开更多
目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织...目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织和细胞系(PC-3和LNCaP)中CSRP1的表达;将CSRP1基因小干扰RNA转染PC-3细胞,下调细胞中CSRP1的表达,采用Transwell小室试验和划痕试验分别检测PC-3细胞的侵袭和迁移能力,Western blot法检测内皮间质转化(epithelial-mesenchymal transition,EMT)标志物E-cadherin和Vimentin的表达水平。结果CSRP1在前列腺癌组织中呈高表达;与正常前列腺上皮细胞RWPE-1相比,CSRP1在2种不同的前列腺癌细胞系中表达水平显著升高(P<0.05)。干扰CSRP1表达后,PC-3细胞的侵袭和迁移能力显著减弱(P<0.05),且E-cadherin的表达水平显著升高(P<0.05),Vimentin的表达受到抑制(P<0.05)。结论CSRP1在前列腺癌组织和细胞系中表达上调。沉默CSRP1表达后可下调Vimentin,逆转EMT的发生,从而抑制前列腺癌细胞的侵袭转移。CSRP1可能是前列腺癌的治疗靶点和潜在的诊断标志物。展开更多
文摘To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable informat
基金financially supported by the National Natural Science Foundation of China (31171565 and 31371635)
文摘The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied physiological and biochemical processes especially in the stress response of plants. In this study, a total of 23 RB‐GRPs distributed on 10 chromosomes were identified in maize（Zea mays L.）, and they were divided into four subgroups according to their conserved domain architecture. Five pairs of paralogs were identified,while none of them was located on the same chromosomal region, suggesting that segmental duplication is predominant in the duplication events of the RB‐GRPs in maize. Comparative analysis of RB‐GRPs in maize, Arabidopsis（Arabidopsis thaliana L.）, rice（Oryza sativa L.）, and wheat（Triticum aestivum）revealed that two exclusive subgroups were only identified in maize. Expression of eight ZmRB‐GRPs was significantly regulated by at least two kinds of stresses. In addition, cis‐elements predicted in the promoter regions of the ZmRB‐GRPs also indicated that these ZmRB‐GRPs would be involved in stress response of maize. The preliminary genome‐wide analysis of the RB‐GRPs in maize would provide useful information for further study on the function of the ZmRB‐GRPs.
文摘BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previous study has demonstrated that CRP2 can be detected in the embryonic rat inner ear but not in the adult rat inner ear. However, at present, the expression of LIM protein family H members in stem or precursor cells has not been described. OBJECTIVE： To determine the expression and sub-cellular localization of CRP2 in olfactory stem cells. DESIGN, TIME AND SETTING： An experiment with repeated measures was performed in the Laboratory of Otorhinolaryngology, Head and Neck Surgery, Xijing Hospital, the Fourth Military Medical University from February 2008 to April 2008. MATERIALS： Olfactory stem cells, and rabbit-anti-CRP2 polyclonal antibody were prepared and kept in our laboratory. METHODS： Reverse transcription polymerase chain reaction and Western blot analysis were used to detect expression of CRP2 in olfactory stem cells. Immunocytochemistry was also used to localize CRP2 in olfactory stem cells. MAIN OUTCOME MEASURES： The expression and sub-cellular localization of CRP2 in rat olfactory stem cells. RESULTS： CRP2 expression was found in olfactory stem cells, and CRP2 was distributed in both the nucleus and the cytoplasm. CONCLUSION： Confirmation of the expression and distribution of CRP2 in olfactory stem cells.
文摘目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织和细胞系(PC-3和LNCaP)中CSRP1的表达;将CSRP1基因小干扰RNA转染PC-3细胞,下调细胞中CSRP1的表达,采用Transwell小室试验和划痕试验分别检测PC-3细胞的侵袭和迁移能力,Western blot法检测内皮间质转化(epithelial-mesenchymal transition,EMT)标志物E-cadherin和Vimentin的表达水平。结果CSRP1在前列腺癌组织中呈高表达;与正常前列腺上皮细胞RWPE-1相比,CSRP1在2种不同的前列腺癌细胞系中表达水平显著升高(P<0.05)。干扰CSRP1表达后,PC-3细胞的侵袭和迁移能力显著减弱(P<0.05),且E-cadherin的表达水平显著升高(P<0.05),Vimentin的表达受到抑制(P<0.05)。结论CSRP1在前列腺癌组织和细胞系中表达上调。沉默CSRP1表达后可下调Vimentin,逆转EMT的发生,从而抑制前列腺癌细胞的侵袭转移。CSRP1可能是前列腺癌的治疗靶点和潜在的诊断标志物。
