| 研究実績の概要 |
Comprehensive sets of purified recombinant mammalian condensin II mutant complexes have been tested in the Xenopus laevis egg extract system to analyze two major condensin II activities: chromosome association and axis formation. Comparisons between the condensin II holocomplex and different condensin II mutants (i.e., subcomplexes lacking a subunit or complexes with mutations) have revealed a wide range of abnormal chromosomal phenotypes. Particular mutants tested include complexes containing mutations in SMC2 and SMC4 ATPase and in potential DNA binding regions of CAP-H2. Through the analysis of condensin II mutant-mediated phenotypes, the functional role of each condensin II subunit (SMC2, SMC4, CAP-D3, CAP-G2, and CAP-H2) in chromosome association and axis formation has been identified. The results suggest both similarities and differences between condensin II subunits and condensin I subunits in their roles in chromosome assembly. Additionally, analysis of recombinant condensin II mutants in the Xenopus laevis egg extract system has revealed the mechanism of condensin II-binding protein MCPH1, a chromosome condensation regulator mutated in microcephaly patients. A general model for the roles of each condensin II subunit in chromosome assembly has been established.
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| 次年度使用額が生じた理由 |
The expenses for FY2020 were used for constructs and reagents necessary for the purification of recombinant condensin II complexes and the depletion of endogenous condensins in the Xenopus laevis egg extract system. Analysis with the currently purified recombinant complexes in Xenopus laevis egg extract has mostly been completed. The incurring amount for FY2021 will include tools and reagents to establish other biochemical assays to test the recombinant condensin II complexes. Additionally, the expenses will be used for constructs to express and purify new recombinant condensin II mutants to analyze the functions of condensin II subunits in greater detail. Furthermore, the expenses will be used for the expression and purification of condensin I complex to test the condensins together in the Xenopus laevis egg extract system.
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