Analysis and Characterization of the Functions of New Factor Related to Transcriptional Regulation in Plasmodium falciparum
| Project/Area Number |
22790400
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Parasitology (including Sanitary zoology)
|
|---|
| Research Institution | Research Institute, International Medical Center of Japan |
|---|
Principal Investigator |
YASUDA Kanako (KOMAKI Kanako) 独立行政法人国立国際医療研究センター (50415551)
|
|---|
| Project Period (FY) |
2010 – 2011
|
| Project Status |
Completed (Fiscal Year 2011)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
|---|
| Keywords | マラリア / 転写 / 転写因子 / 細胞周期 / 転写制御 / 転写調節因子 |
|---|
| Research Abstract |
The detailed mechanisms of transcriptional regulation in malaria parasite, Plasmodium falciparum, remains almost unknown. Genome-wide analyses of transcription-associated proteins have revealed a paucity of putative regulatory transcription factors, suggesting that this parasite has unique regulatory transcription factors distinct from those of other eukaryotes. Previously, to identify unique regulatory transcription factors in P. falciparum, we tried purification and identification of nuclear factor which associates specifically with cis-acting enhancer region of pfl-cys-prx, then, about 20 parasite proteins were identified. In this study, to identify the cis-element-binding factor, 4 candidate proteins, which showed high frequency signal detection in the mass spectrometry, were FLAG-tagged and over-expressed in the parasite cells. These over-expressed proteins were purified from the parasite cells through immunoprecipitation with anti-FLAG antibody, then, enhancer-binding activities
… More
of the recombinant proteins were analyzed by EMSA. As results, one of these proteins revealed sequence-specific enhancer-binding activity. Specific peptide antibody against the new identified factor was produced and used in the supershift assay. The supershift band was observed when the antibody was added in the EMSA reaction with cis-acting enhancer region of pfl-cys-prx and the parasite nuclear extract. These results strongly verified that the new cis-element binding factor was identified(this protein was named as "prx regulatory element binding protein": PREBP). The molecular weight of PREBP estimated from the genome database was 130 KDa, although in the mass-spectrometry, only the peptides corresponding to the central 60 KDa region of PREBP was detected, suggesting that the central region functions in the parasite nuclear. However, the over-expression of the central 60 KDa region of PREBP did not affected to the expression of pfl-cys-prx. The N-terminal and C-terminal region of the 130 KDa coding region of PREBP might be necessary for the function of the central region in the nuclear. In addition, recombinant PREBP protein purified from the cell-free translation system did not show the cis-element binding activity. In contrast, recombinant PREBP protein directly purified from the parasite cells showed the activity, suggesting that some specific modification in the parasite cell is needed for the binding activity. Less
|
Report
(3 results)
Research Products
(9 results)
