INT26891
From wiki-pain
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Sentences Mentioned In
Key: | Protein | Mutation | Event | Anatomy | Negation | Speculation | Pain term | Disease term |
A HeLa cell line stably expressing EGFP-hOGG1-Glu326 was generated and the subcellular localization was examined throughout the cell cycle (Figure 3). | |||||||||||||||
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However, no EGFP-hOGG1-Cys326 could be detected in the soluble chromatin or in the nuclear matrix fractions (Figure 7C, lanes 2 and 4).
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In order to elucidate the cellular role of hOGG1 we have investigated both the expression of hOGG1 and its cellular distribution. | |||||||||||||||
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To determine the distribution of EGFP-hOGG1-Cys326 during mitosis, we created stable cell lines overexpressing EGFP-hOGG1-Cys326. | |||||||||||||||
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In contrast, a recent study in which the promoter of HOGG1 was fused to the firefly luciferase gene carried out in HeLa cells showed that the expression of the reporter gene was not modulated during the cell cycle (71). | |||||||||||||||
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In contrast, a recent study in which the promoter of HOGG1 was fused to the firefly luciferase gene carried out in HeLa cells showed that the expression of the reporter gene was not modulated during the cell cycle (71). | |||||||||||||||
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EGFP-hOGG1 cells were synchronized at the G1/S boundary, released and allowed to enter S-phase before being treated with 0.04 ? | |||||||||||||||
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The cell-cycle-dependent expression of HOGG1 was followed by northern blot hybridization of poly(A)+ RNA isolated from synchronized HaCat cells. | |||||||||||||||
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Cell-cycle-dependent expression of HOGG1 gene | |||||||||||||||
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Immunoblotting with anti-hOGG1 antibody revealed EGFP-hOGG1-Cys326 to be present only in the cytoplasmic fraction (Figure 7C, lane 1). | |||||||||||||||
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Transient transfection of HeLa cells with EGFP-hOGG1-Ala326 showed the distribution to be identical to that of EGFP-hOGG1-Cys326 indicating that the presence of the serine residue is important for the correct localization of hOGG1 to the nucleoli (data not shown).
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However, endogenous hOGG1 is barely detected using the antibodies that are presently available [(23) and L. | |||||||||||||||
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We transiently co-transfected HeLa S3 cells with EYFP-hOGG1 and ECFP-PCNA and followed the cells for 20 h. | |||||||||||||||
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Overexpression of EGFP and EGFP-hOGG1 in the stable cell lines was examined in nuclear extracts by western blot using anti-OGG1 and anti-GFP specific antibodies. | |||||||||||||||
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Site-directed mutagenesis of hOGG1 | |||||||||||||||
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The associations with interphasic chromatin and nuclear matrix, and with mitotic condensed chromosomes are disrupted in EGFP-hOGG1-Cys326 | |||||||||||||||
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In this study, we investigated the subcellular localization and expression of hOGG1 during the cell cycle. | |||||||||||||||
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Northern blot hybridization of synchronized HaCat cells showed a cell-cycle-dependent mRNA expression of hOGG1 with increased transcription of the major nuclear and mitochondrial isoforms during S/G2 (Figure 1). | |||||||||||||||
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By using a cell line constitutively expressing hOGG1 fused to enhanced green fluorescence protein (EGFP), we observed a dynamic relocalization of EGFP-hOGG1 to the nucleoli during the S-phase of the cell cycle, and this localization was shown to be linked to transcription. | |||||||||||||||
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To detect EGFP and EGFP-hOGG1 (and the mutants) the membrane was probed with a polyclonal anti-GFP antibody (ab290-50; Abcam), and to detect EGFP-hOGG1 (and the mutants) a monoclonal anti-human OGG1 antibody (clone 7E2; IBL) was used. | |||||||||||||||
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