2012 Fiscal Year Final Research Report
Development of orthogonal combinations of artificial DNA methyltrasferases and their substrates for study of heritable patterns of DNA methylation.
| Project/Area Number |
23710251
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Living organism molecular science
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
NOMURA Wataru 東京医科歯科大学, 生体材料工学研究所, 准教授 (80463909)
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| Project Period (FY) |
2011 – 2012
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| Keywords | 生体分子の化学修飾 |
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| Research Abstract |
Covalent modification of DNA, such as cytosine methylation, can induce heritable gene silencing. If epigenetic modifications can be specifically targeted, new approaches to transcriptional therapy should result. To address this challenge we have constructed methyltransferases that would act only at a desired site by adapting the sequence-enabled assembly strategy. This is the first successful application of the sequence-enable enzyme reassembly approach in vivo. In this study, to determine the functions of split protein domains in DNA binding and methylation, the split domains were expressed and purified separately. Utilizing these domains, DNA binding analyses were performed. The results indicate cooperative binding of the domains to the specific DNA targets. This interaction between the domains shows a direct evidence of assembly on the target sequence of split domains. The complementary protein assays have been shown their usefulness in dissection of protein interaction in mammalian cells. However, a few of direct approach to evaluate kinetics of interaction of split domains have been performed. Moreover, to expand the targetable DNA sequences on genomic DNA, several zinc finger domains were constructed. These domains showed DNA binding on endogenous targets. To perform efficient DNA methylation in mammalian cells, the expression of methyltransferase would be a key factor. Thus, the codon usage of methyltransferase was optimized. The expression of methyltransferase was greatly increased. The present results would expand the knowledge in the design of split protein domains.
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[Journal Article] Low-molecular-weight CXCR4 ligands with variable spacers.2013
Author(s)
Narumi T, Aikawa H, Tanaka T, Hashimoto C, Ohashi N, Nomura W, Kobayakawa T, Takano H, Hirota Y, Murakami T, Yamamoto N, Tamamura H.
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Journal Title
ChemMedChem
Volume: 8
Pages: 118-124
DOI
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[Journal Article] Pharmacophore-based small molecule CXCR4 ligands.2012
Author(s)
Narumi T, Tanaka T, Hashimoto C, Nomura W, Aikawa H, Sohma A, Itotani K, Kawamata M, Murakami T, Yamamoto N, Tamamura H.
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Journal Title
Bioorg. Med. Chem. Lett.
Volume: 22
Pages: 4169-4172
DOI
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[Journal Article] Conjugation of Cell-penetrating Peptides Leads to Identification of Anti-HIV Peptides from Matrix Proteins.2012
Author(s)
Narumi T, Komoriya M, Hashimoto C, Wu H, Nomura W, Suzuki S, Tanaka T, Chiba J, Yamamoto N, Murakami T, and Tamamura H.
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Journal Title
Bioorg. Med. Chem.
Volume: 20
Pages: 1468-1474
DOI
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[Journal Article] Azamacrocyclic Metal Complexes as CXCR4 Antagonists.2011
Author(s)
Tanaka T, Narumi T, Ozaki T, Sohma A, Ohashi N, Hashimoto C, Itotani K, Nomura W, Murakami T, Yamamoto N, and Tamamura H.
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Journal Title
ChemMedChem
Volume: 6
Pages: 834-839
DOI
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[Journal Article] Synthetic C34 Trimer of HIV-1 gp41 Shows Significant Increase of Inhibition Potency.
Author(s)
Nomura W, Hashimoto C, Ohya A, Miyauchi K, Urano E, Tanaka T, Narumi T, Nakahara T, Komano JA, Yamamoto N, and Tamamura H.
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Journal Title
ChemMedChem
Volume: 7
Pages: 205-208
DOI
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