2023 Fiscal Year Research-status Report
キネシン-輸送基質相互作用特異性の解明と新規ウイルス感染阻害アプローチ
| Project/Area Number |
23K06575
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| Research Institution | National Institute of Infectious Diseases |
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Principal Investigator |
アリ フセインハッサン 国立感染症研究所, ウイルス第二部, 主任研究官 (00523515)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Keywords | Kinesins / Specificity / Substrate |
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| Outline of Annual Research Achievements |
We previously identified a poly Lysine motif at the C-terminal part of NTCP to be the binding site for KIF4A. In 2023, we further performed substitution mutation, and identified 2 amino acids to be essential for this interaction by co-immunoprecipitation assays. KIF4 is reported to bind with ITGB1 and transport it to cell surface. We identified a similar poly-lysine motif in ITGB1; we performed substitution mutation of the similar amino acids that were important for the interaction between KIF4A and NTCP; and found that it suppressed the interaction of ITGB1 with KIF4 by immunoprecipitation assays. These data confirmed the sequence in KIF4 substrates that is required for its identification, interaction, and transport by KIF4.
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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
As planed in the research proposal, in 2023 we identified the amino acids required for the interaction of KIF4A with its substrates. We confirmed the importance of these amino acids in 2 different KIF4A substrates and proved that substitution of these amino acids reduced the surface transport of these proteins.
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| Strategy for Future Research Activity |
We previously showed that the interaction of KIF4A and its substrate is indirect. As planed in the submitted project, in FY2024 we are planning to identify the adaptor protein/s which are required for this interaction. Using the wild type C-terminal sequence of NTCP and ITCB1 together with the mutant sequence carrying substitution in only the 2 important amino acids identified in 2023, we will perform co-immunoprecipitation assay, followed by mass spectrometry to identify the adaptor protein which will bind to WT but not mutant sequence. We will then analyze the effect of this protein on KIF4A/NTCP interaction.
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