1997 Fiscal Year Final Research Report Summary
Interaction of protcin and DNA by AFM (Analysis of the bacterial stress responsive factors binding to DNA by Atomic Force Microscopy).
| Project/Area Number |
09044254
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | University of Tsukuba |
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Principal Investigator |
OHTA Toshiko College of Medical Technology and Nursing Professor, 医療技術短期大学部, 教授 (40233134)
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| Co-Investigator(Kenkyū-buntansha) |
PATEL Smia.S. Ohio State University Biochemistry Asociate Professor, 生物科学科生化学研究室, 助教授
HAI Tsonwin Ohio State University Neuroscience Center Asociate Professor, 神経生物工学センター, 助教授
KURAZONO Hisao University of Tsukuba Microbiology Asistant Professor, 基礎医学系, 講師 (90186487)
HAYASHI Hideo University of Tsukuba Microbiology Professor, 基礎医学系, 教授 (40033203)
TAKEYASU Kunio Kyoto University Integrated Human Studies Professor, 人間科学部, 教授 (40135695)
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| Project Period (FY) |
1997
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| Keywords | Atomic Force Microscopy / Staphylococcus aureus / Heat shock protein / Stress response / Regulator / CIRCE element |
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| Research Abstract |
(1) Disruption and complcmentation of hrc37 of the HSP70 operon in Staphylococcus aureus showed that the transcripts from those CIRCE (Controlling lnvertcd Repeat of Chaperon Expression) depcndcnt promoters were repressed by Hrc37 over-products ; however transcripts from the CIRCE independent promoters were not affected in this case. (submitted)
(2) Furthermore, gel retardation assay and Atomic Force Microscopy (AFM) with a specific CIRCE probe revealed that the Hrc37 could be involved in CIRCE dependent repression of heat induction.
(3) Sigma factor sigB-knockout mutant revealed that SigB enhanced expression of alkaline responsible asp genes, pathogenic factor-controlled gene, sar, and carotenoid-producing gcncs, crtM/N at stationary phase. The SigB recognized a specific sequence, GGGTAT in the promoters of these genes.
(4) Using differential hybridization analysis, one of the S.aureus genomic library were positively induced by heat, cadmium, mercury and zinc insults. DNA sequencing and
… More
Northern analysis revealed that 1.4kb transcript containing two open reading frames (czrA and czrB) as induced by heat and metal insults. The CzrA may play a repressor role in order to autoregulate its own operon. (submitted)
(5) We found a novel 35-kDa membrane protein induced especially by antibiotics that affect S.aureus cell wall. Analysis of SDS-PAGE revealed that a 35-kDa protein band was evidently enhanced after 30min in the presence of beta-lactams. The gene was cloned by differential hybridization with mRNAs enhanced in the prcsence of beta-lactams. The enzymatic activities of the gene were analyzed to make clear its functional role.
(6) A novel transcriptional factor ATF4 was purified and characterized biochemically. When ATF4 was reacted with CREB protein, the complex bound an element, which controlled Gadd153. (published)
(7) Structural analyzes of membrane proteins and nucleic acids by atomic force microscopy (AFM) have demonstrated that channel-like structures exist in both ion channels and ion pumps ; and that distinct higher-order structures such as stem-loop can be formed in the promoter regions of genes. (submitted) Less
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[Publications] Yoshida T., Yohda M., Iida T., Maruyama T., Taguchi H., Yoshida M., Yazaki K., Ohta T., Endo I., and Kagawa Y.: "Molecular cloning and expression of group II chaperonin from the hyperthermophilicarchaeum, Thermococcus strain KS-1 : the homooligomeric complexes ofrecombinant alpha and betasubunits have chaperonin activity." J.Biol.Chem.271. 22017-22021 (1997)
Description
「研究成果報告書概要(欧文)」より
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