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Analysis of structural change of XRCC4 by pseudo-phosphorylation using VUV-CD and SAXS
https://repo.qst.go.jp/records/85311
https://repo.qst.go.jp/records/853117825c8d9-dfcd-4536-bfdd-611658ca9ee3
| Item type | 会議発表用資料 / Presentation(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2022-03-16 | |||||
| タイトル | ||||||
| タイトル | Analysis of structural change of XRCC4 by pseudo-phosphorylation using VUV-CD and SAXS | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_c94f | |||||
| 資源タイプ | conference object | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Kai, Nishikubo
× Kai, Nishikubo× 長谷川真保× Yudai, Izumi× Kentaro, Fujii× 松尾光一× 松本義久× Akinari, Yokoya× Kai, Nishikubo× Yudai, Izumi× Kentaro, Fujii× Akinari, Yokoya |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | XRCC4 is a key protein to repair severe DNA damage of double strand breaks (1). The XRCC4 dimer has been reported to form a multimer scaffold with another repair protein, the XRCC4-like factor (XLF). Enzymatically induced phosphorylation at several amino acid residues in XRCC4, such as serine (2), might cause a local change of statically electric charge, resulting conformational alteration to activate the protein. However, the correlation between the structural change and activation has not been understood yet. Full-length XRCC4 was not able to be crystallized because of its intrinsically disordered C-terminal region including several phosphorylation sites. Previously, we have applied circular dichroism (CD) spectral analysis for the full length of wild type XRCC4 (denoted as WT) in an aqueous solution and reported its characteristic secondary structure (3). In the present study, we focused further the structural change by phosphorylation. We prepared a dimer and multimers of WT XRCC4 as well as mutated ones in which serine residues were substituted to an aspartic acid (denoted as S260D and S327D) to mimic phosphorylation in terms of statically electric charge. Spectra of XRCC4 dimer proteins were similar irrespective of phosphorylation. On the other hand, spectra of multimers were significantly different between WT and mutated proteins (FIGURE 1). The composition ratios of the secondary structures were calculated for each sample using a software, SELCON3 (TABLE 1). The results indicate that the multimerization and pseudo-phosphorylation increase β-strands. Small angle X-ray scattering (SAXS) at BL6A at the Photon Factory in KEK was also applied to further obtain structural information of XRCC4. The results suggest that the XRCC4S260D dimer has a slightly larger radius of inertia (Rg) than WT (TABLE 2), which may correspond to the slight increase in β-strands shown in the CD analysis. The β-strand formation by the phosphorylation at the C-terminal might stabilize the NHEJ machinery after DSB repair completion. |
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| 会議概要（会議名, 開催地, 会期, 主催者等） | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | The 26th Hiroshima International Symposium on Synchrotron Radiation | |||||
| 発表年月日 | ||||||
| 日付 | 2022-03-10 | |||||
| 日付タイプ | Issued | |||||
