リン酸化ユビキチンを手がかりにマイトファジーの全貌にせまる

2016 Fiscal Year Annual Research Report

• Project/Area Number: 16F15387
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: 田中 啓二 公益財団法人東京都医学総合研究所, 生体分子先端研究分野, 所長 (10108871)
• Co-Investigator(Kenkyū-buntansha): QUELICONI BRUNO 公益財団法人東京都医学総合研究所, 生体分子先端研究分野, 外国人特別研究員
• Project Period (FY): 2016-04-22 – 2018-03-31
• Keywords: Ubiquitin / Mitochondria / DJ-1 / Parkinson / TAT

Outline of Annual Research Achievements

We produced Ubiquitin and di-ubiquitin, with or without TAT peptide at the N-terminal region. We used these recombinant proteins with several different concentration and incubation time to test permeability. We then checked cellular penetrance using immunofluorescence and comparing staining with or without pre-permeabilization but we were unable to detect the presence of TAT-Ubiquitin inside the cells. We also were unsuccessful in detecting TAT-ubiquitin penetration when incubated in the presence of proteasome inhibitors.
As the technique is not working and the question we aimed to answer were published using a different technical approach we moved to study DJ-1, a protein related to Parkinson and mitochondria. We already have interesting results and we are preparing a paper

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

We followed our original planning, with success, until the data showed that the continuity would not benefit the scientific community and would be a bad of the grant money. Our current objective is producing an interesting set of data with witch we aim to publish at least 2 papers, with one already in preparation stage.
Considering the problems with the first set of objective and that we already worked to collaborate in 2 papers and have two more with first authorship on track. Therefore, we believe that the project is evolution is going as expected.

Strategy for Future Research Activity

We are now finishing the figures to one publication describing the reasons that promote DJ-1 mutants to localize in mitochondria.
We are also working on the characterization of how DJ-1 affects mitochondrial metabolism, gene expression and protein stability at the same time. We have already characterized new substrates for DJ-1 (paper under revision) and we have data showing how these substrates can affect the different systems inside the cell.