Analysis of mouse Rad54 expression and its implications for homologous recombination

Homologous recombination is one of the major pathways for repair of DNA double-strand breaks (DSBs). Important proteins in this pathway are Rad51 and Rad54. Rad51 forms a nucleoprotein filament on single-stranded DNA (ssDNA) that mediates pairing with and strand invasion of homologous duplex DNA with the assist of Rad54. We estimated that the nucleus of a mouse embryonic stem (ES) cells contains on average 4.7x10(5) Rad51 and 2.4x10(5) Rad54 molecules. Furthermore, we showed that the amount of Rad54 was subject to cell cycle regulation. We discuss our results with respect to two models that describe how Rad54 stimulates Rad51-mediated DNA strand invasion. The models differ in whether Rad54 functions locally or globally. In the first model, Rad54 acts in cis relative to the site of strand invasion. Rad54 coats the Rad51 nucleoprotein filament in stoichiometric amounts and binds to the target duplex DNA at the site that is homologous to the ssDNA in the Rad51 nucleoprotein filament. Subsequently, it promotes duplex DNA unwinding. In the second model, Rad54 acts in trans relative to the site of strand invasion. Rad54 binds duplex DNA distant from the site that will be unwound. Translocation of Rad54 along the duplex DNA increases superhelical stress thereby promoting duplex DNA unwinding.     

Identification of candidate genes and mutations in QTL regions for immune responses in chicken

There are two categories of immune responses – innate and adaptive immunity – both having polygenic backgrounds and a significant environmental component. In our study, adaptive immunity was represented by the specific antibody response toward keyhole limpet hemocyanin (KLH); innate immunity was represented by natural antibodies toward lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Defining genetic bases of immune responses leads from defining quantitative trait loci (QTL) toward a single mutation responsible for variation in the phenotypic trait. The goal of the reported study was to define candidate genes and mutations for the immune traits of interest in chicken by performing an association study of SNPs located in candidate genes defined in QTL regions. Candidate genes and SNPs in QTL regions were selected in silico. SNP association was based on a custom SNP panel, GoldenGate genotyping assay (Illumina) and two statistical models: random mixed model and CAR score. The most significant SNP for immune response toward KLH was located in the JMJD6 gene located on GGA18. Four SNPs in candidate genes FOXJ1 (GGA18), EPHB1 (GGA9), PTGER4 (GGAZ) and PRKCB (GGA14) showed association with natural antibodies for LPS. A single SNP in ITGB4 (GGA18) was associated with natural antibodies for LTA. All associated SNPs mentioned above showed additive effects.     

Role of mammalian Rad54 in telomere length maintenance

The homologous recombination (HR) DNA repair pathway participates in telomere length maintenance in yeast but its putative role at mammalian telomeres is unknown. Mammalian Rad54 is part of the HR machinery, and Rad54-deficient mice show a reduced HR capability. Here, we show that Rad54-deficient mice also show significantly shorter telomeres than wild-type controls, indicating that Rad54 activity plays an essential role in telomere length maintenance in mammals. Rad54 deficiency also resulted in an increased frequency of end-to-end chromosome fusions involving telomeres compared to the controls, suggesting a putative role of Rad54 in telomere capping. Finally, the study of mice doubly deficient for Rad54 and DNA-PKcs showed that telomere fusions due to DNA-PKcs deficiency were not rescued in the absence of Rad54, suggesting that they are not mediated by Rad54 activity.     

云南含笑天然居群的表型多样性分析

为揭示云南含笑天然居群表型变异程度和变异规律,以云南昆明地区天然分布的云南含笑为研究对象,调查了6个居群180个单株的14个表型性状,采用巢式方差分析、变异系数、相关分析、Shannon-Weaver多样性指数分析和聚类分析等方法,分析了居群间和居群内表型多样性.结果表明:(1)云南含笑表型性状在居群间和居群内存在极其丰富的多样性,14个表型性状平均表型分化系数(24.38％)小于居群内变异(75.62％),居群内变异是表型变异的主要来源;14个表型性状的变异系数(CV)在16.20％～60.11％之间,表明云南含笑居群内表型性状离散程度较高.(2)对云南含笑各居群的Shannon-Weaver指数分析表明,云南含笑各居群具有丰富的多样性,总体表型多样性指数为1.772.(3)利用居群间欧氏距离进行的UPGMA聚类分析结果表明,云南含笑6个天然居群可以聚为3类,而且表型性状并没有严格依地理距离而聚类.     

桂东不同林龄马尾松人工林的生物量及其分配特征

根据5a、15a、21a、32a、60a生的5个不同林龄的15块1 000m2样地(3次重复)调查资料,利用21株不同年龄和径阶的马尾松样木数据,建立以胸径(D)为单变量的生物量回归方程.采用样木回归分析法(乔木层)和样方收获法(灌木层、草本层、地上凋落物)获取不同林龄马尾松人工林的生物量,并分析了其组成、分配特征及不同林龄生物量的变化趋势.结果表明:(1)林分的总生物量随林龄而增加,5a、15a、21a、32a和60a生马尾松人工林生物量分别为15.03、125.93、183.51、191.53、405.31 Mg/hm2,其中活体植物占75.01％～94.19％,地上凋落物占0.86％～24.99％.(2)层次分配方面乔木层占绝对优势,占90.20％～98.35％,且随林龄的增加而增大,其次为地上凋落物,占0.86％～24.99％;草本层和灌木层生物量较小,分别占0.47％～34.85％和0.32％～27.00％,均随林龄的增加呈递减趋势.(3)乔木层器官分配以干所占比例最高,占49.93％～83.10％,且随林龄而增加;根相对比较稳定,占6.97％～12.82％;枝、叶分别占11.75％～14.83％、1.33％～23.65％,均随林龄增大而下降.灌木层器官分配除幼龄林为根＞枝＞叶,其余的均呈枝＞根＞叶的趋势.草本层中龄林和近熟林生物量地下＞地上,其他林龄生物量地上＞地下.(4)各林龄凋落物生物量在3.48～6.68Mg/hm2,规律性不强.(5)马尾松人工林乔木层各器官及林分生物量具有良好的优化增长模型,其32a生林分生物量高于同林龄的楠木人工林,低于热带雨林,是一种速生丰产、固碳潜力大的优良造林树种.     

The Wavelet-Based Cluster Analysis for Temporal Gene Expression Data

A variety of high-throughput methods have made it possible to generate detailed temporal expression data for a single gene or large numbers of genes. Common methods for analysis of these large data sets can be problematic. One challenge is the comparison of temporal expression data obtained from different growth conditions where the patterns of expression may be shifted in time. We propose the use of wavelet analysis to transform the data obtained under different growth conditions to permit comparison of expression patterns from experiments that have time shifts or delays. We demonstrate this approach using detailed temporal data for a single bacterial gene obtained under 72 different growth conditions. This general strategy can be applied in the analysis of data sets of thousands of genes under different conditions.[1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29]     

Differential contributions of mammalian Rad54 paralogs to recombination, DNA damage repair, and meiosis

Homologous recombination is a versatile DNA damage repair pathway requiring Rad51 and Rad54. Here we show that a mammalian Rad54 paralog, Rad54B, displays physical and functional interactions with Rad51 and DNA that are similar to those of Rad54. While ablation of Rad54 in mouse embryonic stem (ES) cells leads to a mild reduction in homologous recombination efficiency, the absence of Rad54B has little effect. However, the absence of both Rad54 and Rad54B dramatically reduces homologous recombination efficiency. Furthermore, we show that Rad54B protects ES cells from ionizing radiation and the interstrand DNA cross-linking agent mitomycin C. Interestingly, at the ES cell level the paralogs do not display an additive or synergic interaction with respect to mitomycin C sensitivity, yet animals lacking both Rad54 and Rad54B are dramatically sensitized to mitomycin C compared to either single mutant. This suggests that the paralogs possibly function in a tissue-specific manner. Finally, we show that Rad54, but not Rad54B, is needed for a normal distribution of Rad51 on meiotic chromosomes. Thus, even though the paralogs have similar biochemical properties, genetic analysis in mice uncovered their nonoverlapping roles.