2016 Fiscal Year Annual Research Report
Mechanisms of anhydrobiosis-associated gene regulation in the cells and tissues of chironomid Polypedilum vanderplanki.
| Project/Area Number |
15H05622
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
グセフ オレグ 国立研究開発法人理化学研究所, 予防医療・診断技術開発プログラム, マネージャー (30711999)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Keywords | anhydrobiosis / CAGE / PIMT / LEA proteins / HSFT |
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| Outline of Annual Research Achievements |
At this stage of the project, using CAGE data. we were able to show that, heat shock transcription factor (HSTF) is one of the key regulators of transcriptional activation in response to dehydration in the cells of the anhydrobiotic chironomid.We also confirmed it by RNAi experiment. Mapping the CAGE data updated genome assembly revealed new patterns of transcriptional activity in the tissues of chironomids during anhydrobiosis cycle. Most of the "anhydrobiosis-associated" were activated in dry tissues (75%), with ~ 39% of them were active in various tissues in a similar manner. We found some characteristics associated with tissue-specific activation of this group of genes: globins for for fat body, specific oxidoreductases, methyltransferase, LEA, in the brains.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
Analysis of CAGE signal across the genome of the chironomid was a strong tool to identify tissues-specific activity of the anhydrobiosis-related genes. We determined tissue-specific set of protective genes activity. We also could identify HSTF as one of the key potential regulator of anhydrobiosis regulator, and conform it by in vitro experiments. Thus the progress was greater than we initially expected.
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| Strategy for Future Research Activity |
The final year of the current project will be devoted to functional analysis of the identified regulators of anhydrobiosis. We are going to apply Total-Seq and small RNA analysis of Pv11 cells to identify a pool of non-coding RNA interacting with key anhydrobiotic genes. This year will will also implement protocol of CRISP-Cas9 knock out at least on of the key anhydrobiosis-involved transcription factor gene followed by CAGE analysis to identify the regulatory network associate with anhydrobiosis. Finally, we are going to proceed with phospho-proteomic analysis of Pv11 cells upon desiccation, to link gene expression data and actual protein abundance and proteins phosphorylation patter in the dry cells.
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Research Products
(7 results)
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[Journal Article] A new anhydrobiotic midge from Malawi, Polypedilum pembai sp. nov. (Diptera: Chironomidae), closely related to the desiccation tolerant midge, Polypedilum vanderplanki Hinton2017
Author(s)
Richard Cornette, Nao Yamamoto, Masaru Yamamoto, Tadashi Kobayashi, Ninel A. Petrova5, Oleg Gusev, Sachiko Shimura, Takahiro Kikawada, Dylo Pemba And Takashi Okuda
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Journal Title
Systematic Entomology
Volume: accepted
Pages: N/A
Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
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