2000 Fiscal Year Final Research Report Summary
Analysis of molecular structure of nucleosomes consisting of repetitive DNA sequences and their function in chromatin
| Project/Area Number |
10670114
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | NIIGATA UNIVERSITY |
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Principal Investigator |
MISHIMA Yukio Niigata University, School of Medicine, Associate Professor, 医学部, 助教授 (30166003)
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| Project Period (FY) |
1998 – 2000
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| Keywords | repetitive sequences / GGA repeats / chromosome / reconstituted chromatin / DNA-DNA complex / homopurinesequences / microsatellite / DNase I footprinting |
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| Research Abstract |
We investigated the ability of mononucleosomes reconstituted on DNA fragments containing (GGA/TCC)n repeats for DNA-DNA complex formation and examined the functional aspect of GGA-repeats by isolating genomic DNA fragments containing GGA-repeats and by analyzing their chromosomal localization. The results obtained were as follows.
1. We isolated 19 DNA fragments containing (GGA /TCC)n repeats from mouse genome by triplex affinity capture.
2. 19 clones of them were mapped to different locations of 13 chromosomes.
3. Almost half (9/19) of the isolated DNA fragments containing (GGA/TCC)n repeats were enriched to heterochromatin fraction and the remains were equally distributed to hetero- and eu-chromatin fractions. Neither of the clones examined was enriched to euchromatin fraction.
4. The results from gel mobility shift assay and DNase I protection mapping indicated that genomic DNA fragments containing GGA-repeats formed DNA- DNA complexes in a sequence-dependent manner.
5. The results from the analysis of DNA-DNA complex formation of mononucleosomes reconstituted on various fragments of plasmid DNAs which contained different repeat-number of GGA-repeats or different flanking sequences carrying at either or both sites of GGA-repeats indicated that their complexes formations though GGA-repeats were required for both GGA-repeat number and homologous flanking sequences at both sites of GGA-repeats.
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