2019 Fiscal Year Annual Research Report
Genome organization as a source of chromosomal instability in cancer.
| Project/Area Number |
19H03208
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| Research Institution | Kyoto University |
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Principal Investigator |
Canela Andres 京都大学, 白眉センター, 特定准教授 (90837585)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | TOPOISOMERASE2 / DNA 二重鎖切断 / ゲノム安定性 / 染色体転座 / 発かん |
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| Outline of Annual Research Achievements |
During the fiscal year 2019:
1. Role of TOP2 in chromosomal organization
1.1. Analysis of genome organization in the absence of TOP2. I generated cell lines that are deficient for TOP2 that I will use to characterize the role of TOP2 in genome organization. 1.3. Identification of TOP2β interactions with cohesin components by co-immunoprecipitation. I found by co-immunoprecipitation what cohesin components interact with TOP2β and how this interaction is mediated.
2. Role of TOP2 promoting oncogenic translocations
2.1. Determine the kinetics of generation and repair of TOP2cc. With the modifications in the method END-seq, I was able study how TOP2cc are processed in TOP2-free DSBs, determine the kinetics across the genome and identify factors that modulate this process. I found that although TOP2 binding and cleavage complex (TOP2cc) formation is dependent on cohesin, the conversion of TOP2cc into protein-free DSBs is facilitated by transcription. 2.2 Evaluate of the mechanisms of processing and repair of TOP2cc. I found that transcription is mediating the conversion TOP2cc into protein-free DSBs by proteasomal degradation and TDP2 phosphodiesterase activity. 2.3 Investigate how etoposide-dependent DSBs can lead to chromosomal translocations. I found that transcription promotes chromosomal translocation at TOP2cc mainly by accelerating the conversion of TOP2ccs to DSBs, but has less of an impact on stimulating fusions once DNA ends are protein free.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
During the fiscal year 2019:
1. Role of TOP2 in chromosomal organization. I have made significative advances in this aim by accomplishing the points originally proposed.
2. Role of TOP2 promoting oncogenic translocations. I have made extensive advances in this aim. My results revealed the importance of transcription and chromosomal architecture on the initiation, processing, and repair of Topoisomerase2 breaks. This work was published in the fiscal year 2019: Canela A, Maman Y, Huang SN, Wutz G, Tang W, Zagnoli-Vieira G, Callen E, Wong N, Day A, Peters JM, Caldecott KW, Pommier Y, Nussenzweig A. Topoisomerase II-Induced Chromosome Breakage and Translocation Is Determined by Chromosome Architecture and Transcriptional Activity. Mol Cell 75, 252-266 (2019)
10.1016/j.molcel.2019.04.030
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| Strategy for Future Research Activity |
During the fiscal year 2020:
1. Role of TOP2 in chromosomal organization
1.1. Analysis of genome organization in the absence of TOP2. I will use the cells lines generated for TOP2β and TOP2α to analyze the role of TOP2 in genome organization.
1.2. I will monitor how TOP2 is recruited to the loop anchors.
1.3. Measure and characterize the torsional stress across the genome produced by cohesin advancement.
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Research Products
(8 results)
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[Journal Article] 53BP1 Enforces Distinct Pre- and Post-resection Blocks on Homologous Recombination.2020
Author(s)
Callen E, Zong D, Wu W, Wong N, Stanlie A, Ishikawa M, Pavani R, Dumitrache LC, Byrum AK, Mendez-Dorantes C, Martinez P, Canela A, Maman Y, Day A, Kruhlak MJ, Blasco MA, Stark JM, Mosammaparast N, McKinnon PJ, Nussenzweig A
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Journal Title
Molecular Cell
Volume: 77
Pages: 26-38
DOI
Peer Reviewed / Int'l Joint Research
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[Journal Article] Topoisomerase II-Induced Chromosome Breakage and Translocation Is Determined by Chromosome Architecture and Transcriptional Activity.2019
Author(s)
Canela A, Maman Y, Huang SN, Wutz G, Tang W, Zagnoli-Vieira G, Callen E, Wong N, Day A, Peters JM, Caldecott KW, Pommier Y, Nussenzweig A
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Journal Title
Molecular Cell
Volume: 75
Pages: 252-266
DOI
Peer Reviewed / Int'l Joint Research
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