Escherichia coli single-strand binding protein stabilizes specific denatured sites in superhelical DNA

Escherichia coli single-strand binding protein relaxes supercoiled DNA molecules containing the Drosophila melanogaster histone gene repeat unit, by stabilizing denaturation bubbles that map near the boundaries of the genes, at sites that in native chromatin have been shown to be hypersensitive to nucleases. A similar process may contribute to the propagation of such hypersensitive sites after their induction on the activation of gene expression.     

A highly basic histone H4 domain bound to the sharply bent region of nucleosomal DNA

A nucleosomal core particle is composed of two each of histones H2A, H2B, H3 and H4 located inside the particle with approximately 47 base pairs (bp) of DNA wrapped around the octamer in about 1.8 turns of a left-handed superhelix. The path of the superhelix is not smooth; the DNA is sharply bent, or kinked, at positions symmetrically disposed at a distance of about one and four double-helical turns in both directions from the nucleosomal dyad axis (designated as sites +/- 1 and +/- 4 respectively). This non-uniform bending is considered archetypal to other DNA-protein complexes, but its mechanism is not clear (reviewed in ref. 4). DNA-histone chemical cross-linking within the core particle has revealed strong binding of each of the two histone H4 molecules to DNA at a distance of 1.5 helical turns either side of the nucleosomal dyad axis (sites +/- 1.5). In each of these sites, a single flexible domain of H4 was previously shown to contact three points, at about nucleotides 55 and 65 on one strand and nucleotide 88 on the complementary strand, numbering from the 5' terminus of each 147-base strand; these three locations are closely juxtaposed across the highly compressed minor and major grooves (Fig. 1). Here we report that the amino-acid residue of histone H4 cross-linked at the 1.5 site is histidine-18, embedded in a highly basic cluster Lys-Arg-His-Arg-Lys-Val-Leu-Arg which is probably involved in the sharp bending of the DNA double helix at the +/- 1 sites.     

Identification of a CD4 binding site on the beta 2 domain of HLA-DR molecules.

The CD4 and CD8 molecules are transmembrane glycoproteins expressed by functionally distinct subsets of mature T cells. CD4+ and CD8+ T cells recognize antigens on major histocompatibility complex (MHC) class II-bearing and class I-bearing target cells respectively. The ability of monoclonal antibodies against CD4 and CD8 to block antigen recognition by T cells, as well as cell-cell adhesion assays, indicate that CD4 and CD8 bind to nonpolymorphic determinants of class II or class I MHC. Here we demonstrate that soluble recombinant HLA-DR4 molecules from insect cells and HLA-DR-derived peptides bind to immobilized recombinant soluble CD4. CD4 binds recombinant soluble DR4 heterodimers, as well as the soluble DR4-beta chain alone. Furthermore, two out of twelve DR4-beta peptides could interact specifically with CD4. These findings show that CD4 interacts with a region of MHC class II molecules analogous to a previously identified loop in class I MHC proteins that binds CD8 (refs 8, 9).     

Contributions of a Position Amino Acid Residues to the Conformational Stability of GCN4 Leucine Zipper

Introduction Coiled coils[1] are widespread structures found in many natural proteins, such as structural proteins, transcrip- tion factors, receptor proteins, and enzymes[2,3]. Coils are the dominant structure in many fibrous proteins and the oligomeriza…     

基于IPv4网络的IPv6过渡解决方案

在分析IPv6针对IPv4几个主要问题解决方案的基础上,详细论述了从IPv4到IPv6的两个过渡技术,即双协议栈技术和隧道技术。给出了在实际组网中运用这两种技术实现向IPv6过渡的具体方案。     

6to4路由域非对称路径解决方案研究

6to4隧道是IPv4过渡到IPv6时产生的一种过渡技术,该技术能够解决IPv4网络中的IPv6站点之间的通信和IPv6站点和IPv6骨干网之间的通信.本文通过对6to4路由域中存在的非对称路径问题的研究,讨论了6to4 Relay Anycast机制,并提出了基于路由策略的解决方案.在路由策略解决方案中,设计了新的拓扑结构,在该拓扑结构上实施了静态路由、BGP4 等方法,从而消除了非对称路径.     

Polymorphism in the alpha 3 domain of HLA-A molecules affects binding to CD8

Cytotoxic T lymphocytes (CTL) expressing the CD8 glycoprotein recognize peptide antigens presented by class I major histocompatibility complex (MHC) molecules. This correlation and the absence of CD8 polymorphism led to the hypothesis that CD8 binds to a conserved site of class I MHC molecules. Using a cell-cell binding assay we previously demonstrated specific interaction between human class I MHC (HLA-A,B,C) molecules and CD8. Subsequent analysis of the products of 17 HLA-A,B alleles revealed a natural polymorphism for CD8 binding in the human population. Two molecules, HLA-Aw68.1 and HLA-Aw68.2, which do not bind CD8, have a valine residue at position 245 whereas all other HLA-A,B,C molecules have alanine. Site-directed mutagenesis shows that this single substitution in the alpha 3 domain is responsible for the CD8 binding phenotype and also affects recognition by alloreactive and influenza-specific CTL. Our results indicate that CD8 binds to the alpha 3 domain of class I MHC molecules.     

Thermodynamics of the B-Z transition in superhelical DNA

One of the most exciting events in recent years in molecular biology was the discovery of the left-handed Z form of the DNA double helix. Originally found in linear self-complementary d(GC)x . d(GC)x polymers and oligomers in non-physiological conditions (a rather high salt concentration), it was recently shown to be easily enough adopted in physiological conditions when purine-pyrimidine sequences are inserted into superhelical DNA. From such a system, superhelical DNA carrying an artificial purine-pyrimidine insert, we can obtain data allowing the determination of the energy of the junction between the B and Z stretches, Fj, and the free energy change delta FBZ per base pair (bp). We present here a simple thermodynamic consideration of the B-Z transition in such a system. By applying the results to experimental data we have shown that the thermodynamic parameters for both sequences studied so far (d(GC)x . d(GC)x and d(GT)x . d(AC)x) are similar and equal to Fj = 4-5 kcal per mol per junction and delta FBZ = 0.5 divided by 0.7 kcal per mol per bp.