Crystal structure of thymine DNA glycosylase conjugated to SUMO-1

Members of the small ubiquitin-like modifier (SUMO) family can be covalently attached to the lysine residue of a target protein through an enzymatic pathway similar to that used in ubiquitin conjugation, and are involved in various cellular events that do not rely on degradative signalling via the proteasome or lysosome. However, little is known about the molecular mechanisms of SUMO-modification-induced protein functional transfer. During DNA mismatch repair, SUMO conjugation of the uracil/thymine DNA glycosylase TDG promotes the release of TDG from the abasic (AP) site created after base excision, and coordinates its transfer to AP endonuclease 1, which catalyses the next step in the repair pathway. Here we report the crystal structure of the central region of human TDG conjugated to SUMO-1 at 2.1 A resolution. The structure reveals a helix protruding from the protein surface, which presumably interferes with the product DNA and thus promotes the dissociation of TDG from the DNA molecule. This helix is formed by covalent and non-covalent contacts between TDG and SUMO-1. The non-covalent contacts are also essential for release from the product DNA, as verified by mutagenesis.     

A global heuristically search algorithm for DNA encoding

A new efficient algorithm is developed to design DNA words with equal length for DNA computing. The algorithm uses a global heuristic optimizing search approach and converts constraints to a carry number to accelerate the convergence, which can generate a DNA words set satisfying some thermodynamic and combinatorial constraints. Based on the algorithm, a software for DNA words design is developed.     

Improved taboo search algorithm for designing DNA sequences

The design of DNA sequences is one of the most practical and important research topics in DNA computing. We adopt taboo search algorithm and improve the method for the systematic design of equal-length DNA sequences, which can satisfy certain combinatorial and thermodynamic constraints. Using taboo search algorithm, our method can avoid trapping into local optimization and can find a set of good DNA sequences satisfying required constraints.     

Improved taboo search algorithm for designing DNA sequences

The design of DNA sequences is one of the most practical and important research topics in DNA computing. We adopt taboo search algorithm and improve the method for the systematic design of equal-length DNA sequences, which can satisfy certain combinatorial and thermodynamic constraints. Using taboo search algorithm, our method can avoid trapping into local optimization and can find a set of good DNA sequences satisfying required constraints.     

A global heuristically search algorithm for DNA encoding

A new efficient algorithm is developed to design DNA words with equal length for DNA computing. The algorithm uses a global heuristic optimizing search approach and converts constraints to a carry number to accelerate the convergence, which can generate a DNA words set satisfying some thermodynamic and combinatorial constraints. Based on the algorithm, a software for DNA words design is developed.     

An unprecedented nucleic acid capture mechanism for excision of DNA damage

DNA glycosylases that remove alkylated and deaminated purine nucleobases are essential DNA repair enzymes that protect the genome, and at the same time confound cancer alkylation therapy, by excising cytotoxic N3-methyladenine bases formed by DNA-targeting anticancer compounds. The basis for glycosylase specificity towards N3- and N7-alkylpurines is believed to result from intrinsic instability of the modified bases and not from direct enzyme functional group chemistry. Here we present crystal structures of the recently discovered Bacillus cereus AlkD glycosylase in complex with DNAs containing alkylated, mismatched and abasic nucleotides. Unlike other glycosylases, AlkD captures the extrahelical lesion in a solvent-exposed orientation, providing an illustration for how hydrolysis of N3- and N7-alkylated bases may be facilitated by increased lifetime out of the DNA helix. The structures and supporting biochemical analysis of base flipping and catalysis reveal how the HEAT repeats of AlkD distort the DNA backbone to detect non-Watson-Crick base pairs without duplex intercalation.     

A covalent adduct between the uracil ring and the active site of an aminoacyl tRNA synthetase

A covalent adduct of an aminoacyl tRNA synthetase and uracil nucleoside has been isolated. The enzyme adduct is catalytically inactive; one nucleoside is bound per catalytic site. The release of uridine restores enzyme activity. The nucleoside attaches to a protein segment required for tRNA interaction. The findings add support to concepts of a covalent component for some protein-nucleic acid complexes.     

胸腺嘧啶紫外损伤与自修复机理的计算机模拟

为了研究DNA分子紫外光辐射下光损伤和自修复,采用半经典动力学方法模拟了胸腺嘧啶分子激发与失活过程.模拟脉冲频率为5.0 eV,fwhm=50 fs.模拟发现胸腺嘧啶在紫外辐射下会生成电子激发态,导致分子结构畸变,可能引发突变.由于激发态寿命极短,激发态的胸腺嘧啶分子通过H6原子和C6原子的畸变发生无辐射失活而衰减至基...     

查找算法平均查找长度的计算方法

给出了常用查找算法平均查找长度的计算方法,包括查找成功和查找失败平均查找长度的计算,并通过实例进行了解析,便于学习者学习和掌握查找算法,同时为应用者选择查找算法提供依据。     

垂直搜索在个性化Web搜索中的应用

本文先介绍了个性化Web搜索,然后根据个性化Web搜索提出的要求引出了垂直搜索技术,并探讨了与通用搜索引擎相比较而言．将垂直搜索应用于个性化Web搜索的优越性。最后介绍了垂直搜索中的关键性技术,即信息提取技术,并着重给出了基于网页布局和关键字段的信息提取技术。     

Structure and mechanism of the uracil transporter UraA

The nucleobase/ascorbate transporter (NAT) proteins, also known as nucleobase/cation symporter 2 (NCS2) proteins, are responsible for the uptake of nucleobases in all kingdoms of life and for the transport of vitamin C in mammals. Despite functional characterization of the NAT family members in bacteria, fungi and mammals, detailed structural information remains unavailable. Here we report the crystal structure of a representative NAT protein, the Escherichia coli uracil/H(+) symporter UraA, in complex with uracil at a resolution of 2.8??. UraA has a novel structural fold, with 14 transmembrane segments (TMs) divided into two inverted repeats. A pair of antiparallel β-strands is located between TM3 and TM10 and has an important role in structural organization and substrate recognition. The structure is spatially arranged into a core domain and a gate domain. Uracil, located at the interface between the two domains, is coordinated mainly by residues from the core domain. Structural analysis suggests that alternating access of the substrate may be achieved through conformational changes of the gate domain.     

食品中葡萄糖的酶—比色法分析

用葡萄糖氧化酶和过氧化物酶作酶试剂，苯酚和４氨基安替吡啉作显色剂比色测定食品中的葡萄糖．测定的最大吸收波长为５０５ｎｍ，酶促反应稳定时间４０ｍｉｎ．２４ｈ内重现性好，相对误差小于２％．３２种食品分析确定标准系列范围、称样量．样品测定的不确定度小于０．０９８ｍｇ／ｇ，检出限为０．００３３％．     

一种检测电芬顿体系下DNA损伤的电化学生物传感器的研制

为了检测电芬顿体系下DNA的损伤,先采用石墨烯制备了一种致密的rGO/Fe₃O₄复合材料;再将复合材料和DNA修饰到玻碳电极上,利用电化学还原作用释放游离态Fe²⁺,并加入H₂O₂形成电芬顿体系;最后构建了一种检测电芬顿体系下DNA损伤的新型电化学生物传感器。检测结果表明,检测DNA损伤的最佳时间为30 min,最佳pH值为7.0。     

Enzymatic incorporation of a new base pair into DNA and RNA extends the genetic alphabet.

A new Watson-Crick base pair, with a hydrogen bonding pattern different from that in the A.T and G.C base pairs, is incorporated into duplex DNA and RNA by DNA and RNA polymerases and expands the genetic alphabet from 4 to 6 letters. This expansion could lead to RNAs with greater diversity in functional groups and greater catalytic potential.     

Single-molecule imaging of DNA pairing by RecA reveals a three-dimensional homology search

DNA breaks can be repaired with high fidelity by homologous recombination. A ubiquitous protein that is essential for this DNA template-directed repair is RecA. After resection of broken DNA to produce single-stranded DNA (ssDNA), RecA assembles on this ssDNA into a filament with the unique capacity to search and find DNA sequences in double-stranded DNA (dsDNA) that are homologous to the ssDNA. This homology search is vital to recombinational DNA repair, and results in homologous pairing and exchange of DNA strands. Homologous pairing involves DNA sequence-specific target location by the RecA-ssDNA complex. Despite decades of study, the mechanism of this enigmatic search process remains unknown. RecA is a DNA-dependent ATPase, but ATP hydrolysis is not required for DNA pairing and strand exchange, eliminating active search processes. Using dual optical trapping to manipulate DNA, and single-molecule fluorescence microscopy to image DNA pairing, we demonstrate that both the three-dimensional conformational state of the dsDNA target and the length of the homologous RecA-ssDNA filament have important roles in the homology search. We discovered that as the end-to-end distance of the target dsDNA molecule is increased, constraining the available three-dimensional (3D) conformations of the molecule, the rate of homologous pairing decreases. Conversely, when the length of the ssDNA in the nucleoprotein filament is increased, homology is found faster. We propose a model for the DNA homology search process termed 'intersegmental contact sampling', in which the intrinsic multivalent nature of the RecA nucleoprotein filament is used to search DNA sequence space within 3D domains of DNA, exploiting multiple weak contacts to rapidly search for homology. Our findings highlight the importance of the 3D conformational dynamics of DNA, reveal a previously unknown facet of the homology search, and provide insight into the mechanism of DNA target location by this member of a universal family of proteins.