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Protein and mRNA levels of IgM H- and L-chains artificially and excessively accumulated in HDAC2-deficient DT40 mutants are dramatically reduced through various generations during continuous cultivation
http://hdl.handle.net/10458/6500
http://hdl.handle.net/10458/6500211fb566-1ed7-4183-90a9-997c879ff501
| Item type | その他 / Others(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2020-06-21 | |||||
| タイトル | ||||||
| タイトル | Protein and mRNA levels of IgM H- and L-chains artificially and excessively accumulated in HDAC2-deficient DT40 mutants are dramatically reduced through various generations during continuous cultivation | |||||
| 言語 | en | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | HDAC2-deficient DT40 mutants, continuous cultivation, decreases in IgM H/L chain protein/mRNA levels, changes in mRNA levels of transcription factors, gene targeting techniques, ChIP assay, changes in acetylation levels of K9/H3 surrounding Pax5 5'-upstream chromatin region | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_1843 | |||||
| 資源タイプ | other | |||||
| 著者 |
中山, 雅美
× 中山, 雅美× 中山, 建男 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | We analyzed histone deacetylase-2 （HDAC2）-deficient DT40 mutant cells, HDAC2（-/-）, which were cultivated for varying periods, by two-dimensional polyacrylamide gel electrophoresis （2D-PAGE）, Western blotting, reverse transcription-polymerase chain reaction （RT-PCR） and immuno-electron microscopy. Protein and mRNA levels of IgM H- and L-chains were increased at the early stage of cultivation and thereafter until the later stage reduced to comparable levels as in DT40 cells. The mRNA level of PCAF was increased from the early to later stages, and those of HDAC7 and HDAC9 changed during cultivation. The mRNA level of EBF1 was almost completely decreased at the early stage and thereafter remained unchanged, but that of E2A was slightly increased at the early stage and thereafter remained unchanged. The mRNA level of Pax5, which down-regulates gene expressions of IgM H- and L-chains, was reduced at the early stage and thereafter increased through the middle to later stages in anti-parallel with those of these two immunoglobulin proteins. Bulk acetylation levels of Lys-9, Lys-14, Lys-18, Lys-23 and Lys-27 residues of histone H3 （K9/H3, K14/H3, K18/H3, K23/H3 and K27/H3） were increased during cultivation. Chromatin immuno-precipitation （ChIP） assay suggested that acetylation levels of K9/H3 within some chromatin segments surrounding proximal ~2.0 kb 5'-upstream region of the Pax5 gene were decreased at the early stage in HDAC2（-/-） and thereafter at the later stage increased to comparable levels as in DT40 cells. These results revealed not only that the HDAC2-deficiency induces dramatic increases in mRNA and protein levels of IgM H- and L-chains, and thereafter these accumulated mRNAs and proteins are gradually reduced during cultivation, but also that alterations in gene expressions of IgM H- and L-chains may be coupled with changes in those of Pax5, EBF1, PCAF, HDAC7 and HDAC9. In addition, the changing pattern in acetylation levels of K9/H3 within proximal 5'-upstream chromatin region of the Pax5 gene in HDAC2（-/-） qualitatively agrees with that in the transcription level of the gene. |
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| 言語 | en | |||||
| 内容記述 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | The original edition of the revision is available from following URL: http://opac2.lib.miyazaki-u.ac.jp/webopac/TC10145170 | |||||
| 言語 | en | |||||
| 著者版フラグ | ||||||
| 出版タイプ | AM | |||||
| 出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
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中山, 雅美, 中山, 建男, n.d., Protein and mRNA levels of IgM H- and L-chains artificially and excessively accumulated in HDAC2-deficient DT40 mutants are dramatically reduced through various generations during continuous cultivation.
