HISTONES AND NUCLEOSOMES IN ARCHAEA (ARCHAEBACTERIA)
古细菌(古细菌)中的组蛋白和核小体
基本信息
- 批准号:2907400
- 负责人:
- 金额:$ 17.75万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1995
- 资助国家:美国
- 起止时间:1995-09-01 至 2003-06-30
- 项目状态:已结题
- 来源:
- 关键词:Archaea X ray crystallography biochemical evolution chromatin circular DNA crystallization dimer histones intermolecular interaction microorganism genetics molecular cloning nucleic acid sequence nucleosomes protein protein interaction protein structure function recombinant proteins site directed mutagenesis species difference
项目摘要
We have discovered that the Archaea (formerly Archaebacteria) contain
small DNA binding proteins that have primary sequences, and predicted
secondary structures, in common with the nucleosome core histones. These
archaeal histones bind and compact DNA in vivo and in vitro into
structures which visibly resemble nucleosomes. This discovery, coupled
with the recent recognition that all the basic molecular components of
transcription initiation are conserved in the Archaea and Eukarya
(formerly Eukaryotes) suggests that the structure, and the functions of
nucleosomes in genome compaction and in regulating gene expression, have
evolved from a simpler system that is still retained in the Archaea. The
experiments proposed are to determine if this is correct. We will
determine the 3D structure of HMfA and HMfB, histones A and B from
Methanothermus fervidus, and the molecular stoichiometry and architecture
of the nucleosome-like structures (NLS) that they form. The protein-DNA
interactions that determine where and how these archaeal histones bind and
constrain DNA molecules into NLS will be determined by site-directed
mutagenesis and by the isolation, selection and characterization of
preferred and high affinity DNA binding sites. Nucleosomes are
specifically positioned and re-positioned within chromatin to regulate
eukaryotic gene expression at the level of transcription initiation, and
we will determine if this is also the case for the archaeal NLS. We have
established conditions that turn on and off four very strong archaeal
promoters in vivo, and also procedures to isolate NLS assembled and cross-
linked in situ in these cells under these different conditions. We will
determine where NLS are assembled in vivo, in relationship to the sites at
which these promoters direct transcription initiation, under conditions of
promoter activity and inactivity, by identifying nuclease protected sites
by probe hybridizations, indirect end-labeling and primer extension
procedures. Determining how gene expression signals are accessed and
activated from within chromatin remains one of the most important
questions in biomedical research, and one of the most difficult to address
experimentally. The experiments proposed will not only establish if
histones, nucleosomes and chromatin originated in the Archaea, but also
the extent to which this simpler system provides a valid model for studies
of the structure of the nucleosome, of nucleosome positioning, and of the
mechanisms by which nucleosomes regulate specific gene expression.
我们已经发现,古生菌(原古细菌)含有
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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JOHN NEWTON REEVE其他文献
JOHN NEWTON REEVE的其他文献
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{{ truncateString('JOHN NEWTON REEVE', 18)}}的其他基金
A Model Archaeal System: Genetic Tools and Resources for T. kodakarensis
模型古菌系统:T. kodakarensis 的遗传工具和资源
- 批准号:
8322564 - 财政年份:2011
- 资助金额:
$ 17.75万 - 项目类别:
A Model Archaeal System: Genetic Tools and Resources for T. kodakarensis
模型古菌系统:T. kodakarensis 的遗传工具和资源
- 批准号:
8720014 - 财政年份:2011
- 资助金额:
$ 17.75万 - 项目类别:
A Model Archaeal System: Genetic Tools and Resources for T. kodakarensis
模型古菌系统:T. kodakarensis 的遗传工具和资源
- 批准号:
8150713 - 财政年份:2011
- 资助金额:
$ 17.75万 - 项目类别:
A Model Archaeal System: Genetic Tools and Resources for T. kodakarensis
模型古菌系统:T. kodakarensis 的遗传工具和资源
- 批准号:
8535171 - 财政年份:2011
- 资助金额:
$ 17.75万 - 项目类别:
HISTONES AND NUCLEOSOMES IN ARCHAEA (ARCHAEBACTERIA)
古细菌(古细菌)中的组蛋白和核小体
- 批准号:
2192487 - 财政年份:1995
- 资助金额:
$ 17.75万 - 项目类别:
HISTONES AND NUCLEOSOMES IN ARCHAEA (ARCHAEBACTERIA)
古细菌(古细菌)中的组蛋白和核小体
- 批准号:
6519674 - 财政年份:1995
- 资助金额:
$ 17.75万 - 项目类别:
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