Interactions between the Promoter Regions of Nitrogenase Structural Genes (nifHDK2) and DNA-Binding Proteins from N2- and Ammonium-Grown Cells of the Archaeon Methanosarcina barkeri 227

Yueh-tyng Chien; John D. Helmann; Stephen H. Zinder

首页> 外文期刊>Journal of bacteriology >Interactions between the Promoter Regions of Nitrogenase Structural Genes (nifHDK2) and DNA-Binding Proteins from N2- and Ammonium-Grown Cells of the Archaeon Methanosarcina barkeri 227

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Interactions between the Promoter Regions of Nitrogenase Structural Genes (nifHDK2) and DNA-Binding Proteins from N2- and Ammonium-Grown Cells of the Archaeon Methanosarcina barkeri 227

机译：氮细菌结构基因（nifHDK2）的启动子区域与古细菌Methanosarcina barkeri 227的N2-和铵态生长细胞的DNA结合蛋白之间的相互作用

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Transcription initiation in Archaea (archaebacteria) resembles the eucaryotic process, having been shown to involve TATA box-like promoter regions as well as TATA-binding protein and TFIIB homologs. However, little is known about transcription regulation in archaea. We have previously demonstrated that transcripts ofnifHDK2 genes, encoding Methanosarcina barkerinitrogenase, are present in N₂-grown cells but not in ammonium-grown cells, indicating that nif transcription is regulated by the nitrogen source. In this study, we detected proteins in M. barkeri cell extracts that bind specifically to DNA containing the putative promoter region of nifHDK2. No binding was found when the promoter region was deleted from the DNA. A competition assay showed that the methyl coenzyme M reductase (mcr) promoter region DNA and the nifH2promoter region DNA competed for a common factor(s). There was no binding to the nifH2 promoter region by extracts of ammonium-grown cells, but there was binding by these extracts to promoter regions for mcr genes, which are presumably constitutively expressed. Interestingly, extracts of ammonium-grown cells inhibited binding to the nif promoter region by extracts of N₂-grown cells. Fractionation of extracts of ammonium-grown cells with a heparin-Sepharose column resolved them into a fraction eluting at 0 M NaCl, which inhibited binding by extracts of N₂-grown cells, and a fraction eluting at 0.5 to 0.75 M NaCl, which showed binding to the promoter region. These results are congruent with a model for regulation of nif gene expression in M. barkeri in which a substance present in ammonium-grown cells inhibits DNA binding by a transcription-associated protein or proteins.

机译： Archaea （古细菌）中的转录起始类似于真核生物过程，已被证实涉及TATA盒状启动子区域以及TATA结合蛋白和TFIIB同源物。然而，关于古细菌中的转录调控知之甚少。先前我们已经证明， nifHDK2 基因的转录本，编码甲烷甲烷八叠球菌氮氧化酶，存在于N _{2 生长的细胞中，但不存在于铵生长的细胞中。细胞，表明 nif 转录受氮源调控。在这项研究中，我们检测到 M中的蛋白质。可以与含有 nifHDK2 的假定启动子区域的DNA特异性结合的barkeri 细胞提取物。当启动子区域从DNA中缺失时，未发现结合。竞争测定表明，甲基辅酶M还原酶（ mcr ）启动子区域DNA和 nifH2 启动子区域DNA竞争同一因子。铵生长细胞的提取物不与 nifH2 启动子区域结合，但这些提取物与 mcr 基因的启动子区域结合，这些基因可能是组成型表达的。。有趣的是，铵生长的细胞提取物通过N _{2 生长的细胞提取物抑制了与 nif 启动子区域的结合。用肝素-琼脂糖柱对铵生长的细胞提取物进行分级分离，将其分离为0 M NaCl洗脱的馏分，该馏分抑制了N <2 生长的细胞的提取物的结合，而在0.5 M NaCl洗脱的馏分中至0.75 M NaCl，显示出与启动子区域的结合。这些结果与 M中 nif 基因表达调控模型相吻合。 barkeri ，其中存在于铵生长的细胞中的一种物质通过一种或多种转录相关蛋白抑制DNA结合。}} 展开▼

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《Journal of bacteriology》 |1998年第10期|共6页

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Yueh-tyng Chien; John D. Helmann; Stephen H. Zinder;
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1. Cloning, functional organization, transcript studies, and phylogenetic analysis of the complete nitrogenase structural genes (nifHDK2) and associated genes in the archaeon Methanosarcina barkeri 227. [J] . Y T Chien, S H Zinder Journal of bacteriology . 1996,第1期

机译：对古细菌甲烷八叠球菌227中完整的固氮酶结构基因（nifHDK2）和相关基因的克隆，功能组织，转录本研究和系统发育分析。

2. Analysis of Genes Encoding an Alternative Nitrogenase in the Archaeon Methanosarcina barkeri227 [J] . Yueh-Tyng Chien, Victoria Auerbuch, Andrew D. Brabban, Journal of bacteriology . 2000,第11期

机译：巴氏古生甲烷菌中编码替代性固氮酶的基因分析

3. Cloning, DNA sequencing, and characterization of a nifD-homologous gene from the archaeon Methanosarcina barkeri 227 which resembles nifD1 from the eubacterium Clostridium pasteurianum. [J] . Y T Chien, S H Zinder Journal of bacteriology . 1994,第21期

机译：来自古细菌Methanosarcina barkeri 227的nifD同源基因的克隆，DNA测序和表征，类似于真细菌巴斯德氏梭菌的nifD1。

4. Structural Protein Interactions Predict Kinase-Inhibitor Interactions in Upregulated Pancreas Tumour Genes Expression Data [C] . Gihan Dawelbait, Christian Pilarsky, Yanju Zhang, International Symposium on Computational Life Sciences(CompLife 2005); 20050925-27; Konstanz(DE) . 2005

机译：结构蛋白相互作用预测上调胰腺肿瘤基因表达数据中的激酶抑制剂相互作用。

5. Nitrogen fixation and its regulation in the archaeon Methanosarcina barkeri 227 [D] . Chien, Yueh-tyng. 1996

机译：巴氏甲烷甲烷菌（Methanosarcina barkeri 227）中的固氮及其调控

6. Interactions between the Promoter Regions of Nitrogenase Structural Genes (nifHDK2) and DNA-Binding Proteins from N2- and Ammonium-Grown Cells of the Archaeon Methanosarcina barkeri 227 [O] . Yueh-tyng Chien, John D. Helmann, Stephen H. Zinder 1998

机译：氮细菌结构基因（nifHDK2）的启动子区域与古细菌Methanosarcina barkeri 227的N2-和铵态生长细胞的DNA结合蛋白之间的相互作用

7. Cloning, functional organization, transcript studies, and phylogenetic analysis of the complete nitrogenase structural genes (nifHDK2) and associated genes in the archaeon Methanosarcina barkeri 227. [O] . Chien, Y T, Zinder, S H 1996

机译：对古细菌甲烷八叠球菌227中完整的固氮酶结构基因（nifHDK2）和相关基因的克隆，功能组织，转录本研究和系统发育分析。

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Interactions between the Promoter Regions of Nitrogenase Structural Genes (nifHDK2) and DNA-Binding Proteins from N2- and Ammonium-Grown Cells of the Archaeon Methanosarcina barkeri 227

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