共查询到20条相似文献,搜索用时 31 毫秒
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Nuclear factor III, a novel sequence-specific DNA-binding protein from HeLa cells stimulating adenovirus DNA replication 总被引:76,自引:0,他引:76
Dissection and reconstitution of the adenovirus DNA replication machinery has led to the discovery of two HeLa nuclear proteins which are required in conjunction with three viral proteins. One of these, nuclear factor I (NF-I), recognizes an internal region of the origin between nucleotides 25 and 40 and by binding to one side of the helix stimulates the initiation reaction up to 30-fold. NFI-binding sites have been observed upstream of several cellular genes, such as chicken lysozyme, human IgM and human c-myc, and coincide in most cases with DNase I hypersensitive regions. Here we report the identification of a novel DNA-binding protein from HeLa nuclei, designated NF-III, that recognizes a sequence in the adenovirus origin very close to the NFI-binding site, between nucleotides 36 and 54. This sequence includes the partially conserved nucleotides TATGATAATGAG. NF-III stimulates DNA replication four- to sixfold by increasing the initiation efficiency. Potential cellular binding sites include promoter elements of the histone H2B gene, the human interferon beta gene, the human and mouse immunoglobulin VK and VH genes and the mammal/chicken/Xenopus laevis U1 and U2 small nuclear RNA genes. Furthermore, a subset of the herpes simplex virus immediate early promoter specific TAATGARAT elements is homologous with the adenovirus 2 (Ad-2) NFIII-binding site. 相似文献
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Functionally distinct subsites on a class II major histocompatibility complex molecule 总被引:12,自引:0,他引:12
Mature T lymphocytes are activated by recognition of the combination of foreign protein antigen and membrane products of the major histocompatibility complex (MHC). Studies of peptide antigen binding to detergent-solubilized class II MHC molecules (Ia) have established that peptide-Ia interaction occurs in the absence of the T-cell receptor and varies according to allele-specific features of Ia molecules. The residues of immunogenic peptides thus contribute to two largely independent functions--the control of association with Ia molecules and the determination of the specificity of T-cell receptor binding. Two analogous and potentially independent functional sites have been postulated for Ia molecules--a region that controls binding to peptides and a region that interacts with T-cell receptors. Here we present evidence from functional analysis of recombinant class II molecules that these two postulated functional regions of Ia molecules do exist and can be independently manipulated, consistent with our recent demonstration of the segmental nature of Ia molecule structure-function relationships. 相似文献
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Neutron scattering studies of lac repressor 总被引:3,自引:0,他引:3
The lac repressor, a tetrameric protein of identical subunits [molecular weight (MW) 4 x 38,500], interacts specifically with the lac operator, preventing the expression of the structural genes of the lac operon. The presence of an inducer (such as isopropyl-beta-D-thiogalactoside; IPTG) which binds to the repressor, prevents operator binding by lowering the association constant by a factor of 10(3) (ref. 3). Genetic and biochemical analysis have shown that the major part--if not all--of the binding site for the lac operator is located in the 60 N-terminal residues of the protein. In certain conditions, limited trypsinolysis of the protein yields four N-terminal 'headpieces' each containing 51 or 59 residues, and a tetrameric core with full inducer binding activity. It was shown recently that this headpiece is able to bind nucleic acids, and interacts with the lac operator, giving the same pattern of sensitivity with respect to the methylation of the bases as does the intact repressor. We are studying the interaction of lac repressor with DNA by neutron scattering using contrast variation and discuss here measurements on the protein, its tryptic core and their complexes with IPTG. Our results demonstrate that the headpieces are located far (67 +/- 10 A) from the centre of mass of a somewhat elongated core, and that the inducer does not significantly change the radius of gyration of the protein. 相似文献
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A significant proportion of the genomes of higher plants and vertebrates consists of transposable elements and their derivatives. Autonomous DNA type transposons encode a transposase that enables them to mobilize to a new chromosomal position in the host genome by a cut-and-paste mechanism. As this is potentially mutagenic, the host limits transposition through epigenetic gene silencing and heterochromatin formation. Here we show that a transposase from Arabidopsis thaliana that we named DAYSLEEPER is essential for normal plant growth; it shares several characteristics with the hAT (hobo, Activator, Tam3) family of transposases. DAYSLEEPER was isolated as a factor binding to a motif (Kubox1) present in the upstream region of the Arabidopsis DNA repair gene Ku70. This motif is also present in the upstream regions of many other plant genes. Plants lacking DAYSLEEPER or strongly overexpressing this gene do not develop in a normal manner. Furthermore, DAYSLEEPER overexpression results in the altered expression of many genes. Our data indicate that transposase-like genes can be essential for plant development and can also regulate global gene expression. Thus, transposases can become domesticated by the host to fulfil important cellular functions. 相似文献