The Experts below are selected from a list of 5808 Experts worldwide ranked by ideXlab platform
Jingfang Ju - One of the best experts on this subject based on the ideXlab platform.
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translational control analysis by translationally active rna capture microarray analysis trip chip
Nucleic Acids Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and Polysomeassociated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens.
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abstract 1143 translational control analysis by translationally active rna capture microarray analysis trip chip
Cancer Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil (5-FU) treatment as assessed by real time qRT-PCR analysis. In contrast, the protein expression and Polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1143.
Kenji Kudo - One of the best experts on this subject based on the ideXlab platform.
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translational control analysis by translationally active rna capture microarray analysis trip chip
Nucleic Acids Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and Polysomeassociated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens.
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abstract 1143 translational control analysis by translationally active rna capture microarray analysis trip chip
Cancer Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil (5-FU) treatment as assessed by real time qRT-PCR analysis. In contrast, the protein expression and Polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1143.
Shunsuke Kobayashi - One of the best experts on this subject based on the ideXlab platform.
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hsp60 interacts with yb 1 and affects its Polysome association and subcellular localization
Biochemical and Biophysical Research Communications, 2009Co-Authors: Sachiyo Ohashi, Megumi Atsumi, Shunsuke KobayashiAbstract:YB-1 is a DNA/RNA-binding protein which, in the cytoplasm, associates with Polysomes and regulates translation. However, YB-1 has a novel nuclear localization signal, and its nuclear accumulation is correlated with cancer induction. Here we designated the amino-acid sequence as YB-NLS and demonstrated that YB-NLS is necessary for the nuclear translocation of overexpressed YB-1 in NG108-15 cells. In addition, we found that a heat shock protein, HSP60, binds to YB-NLS in the cytoplasm. Interestingly, when HSP60 expression was repressed, an increase of Polysome-associated YB-1 was observed in heavy-sedimenting fractions on a sucrose gradient. Overexpression of HSP60 resulted in a decrease of YB-1 in the heavy-sedimenting fractions and suppression of YB-NLS activity. Furthermore, the NLS-deleted YB-1 was apparently associated with the heavy-sedimenting Polysomes. These results suggest that HSP60 interacts with YB-1 at the YB-NLS region and acts as a regulator of Polysome association and the subcellular distribution of YB-1.
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expression and Polysome association of yb 1 in various tissues at different stages in the lifespan of mice
Biochimica et Biophysica Acta, 2006Co-Authors: Arisa Miwa, Takashi Higuchi, Shunsuke KobayashiAbstract:Tissue-specific translational regulation is important for gene expression. YB-1 binds to mRNAs to form mRNPs and affects translation. In this study we investigated expression and Polysome association of YB-1 in various tissues at different stages in the lifespan of mice. YB-1 levels decreased markedly with growth in brain, heart and muscle, but increased in the spleen. In lung, kidney and testis, the levels of YB-1 diminished with aging. In liver, no significant change in the level of YB-1 was observed throughout life. We further showed that the distribution pattern of YB-1 on a sucrose gradient differed according to tissue. Moreover, the distribution pattern of YB-1 changed drastically with growth in the liver. In 5-day-old liver, YB-1 was distributed almost exclusively in nonpolysomal fractions, whereas in 4-week-old liver, it was associated with heavy-sedimenting Polysomes, as was the case in 5-day-old brain. Immunohistochemical analysis revealed that YB-1 is mainly a cytoplasmic protein in these tissues. Our results indicate that the expression and Polysome association of YB-1 are regulated with growth or aging in a tissue-specific manner, presumably to control gene expression at the translational level in each tissue.
Yaguang Xi - One of the best experts on this subject based on the ideXlab platform.
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translational control analysis by translationally active rna capture microarray analysis trip chip
Nucleic Acids Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and Polysomeassociated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens.
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abstract 1143 translational control analysis by translationally active rna capture microarray analysis trip chip
Cancer Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil (5-FU) treatment as assessed by real time qRT-PCR analysis. In contrast, the protein expression and Polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1143.
James J Russo - One of the best experts on this subject based on the ideXlab platform.
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translational control analysis by translationally active rna capture microarray analysis trip chip
Nucleic Acids Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and Polysomeassociated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens.
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abstract 1143 translational control analysis by translationally active rna capture microarray analysis trip chip
Cancer Research, 2010Co-Authors: Kenji Kudo, Yaguang Xi, Yuan Wang, Bo Song, Jingyue Ju, James J Russo, Jingfang JuAbstract:We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with Polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with Polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the Polysome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil (5-FU) treatment as assessed by real time qRT-PCR analysis. In contrast, the protein expression and Polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1143.
