Rec-Mediated Recombinational Hot Spot Activity in Bacteriophage Lambda. I. Hot Spot Activity Associated with Spi- Deletions and bio Substitutions

In order to survey the distribution along the bacteriophage lambda chromosome of Rec-mediated recombination events, crosses are performed using conditions which block essentially all DNA synthesis. One parent is density-labeled and carries a genetic marker in the left terminal lambda gene (A), while the other parent is unlabeled and carries a genetic marker in the right terminal lambda gene (R). Both parents are deleted for the lambda recombination genes int and red, together with other recombination-associated genes, by virtue of either (1) a pure deletion or (2) a bio insertion-deletion. The distribution in a cesium density gradient of the resulting A+R+ recombinant phage reflects the chromosomal distribution of the recombination events which gave rise to those phage.Crosses employing either of two different pure deletion phage strains exhibit recombinational hot spot activity located near the right end of the lambda chromosome, between the cI and R genes. This hot spot activity persists when unlimited DNA synthesis is allowed. Crosses employing bio1-substituted phage strains exhibit recombinational hot spot activity located to the right of the middle of the chromosome and to the left of the cI gene. Crosses employing either bio1 or bio69-substituted phage strains indicate that the bio-associated hot spot activity occurs in the presence of DNA synthesis, but is dependent on a functional host recB gene.     

Hot Spots of Recombination in Fission Yeast: Inactivation of the M26 Hot Spot by Deletion of the Ade6 Promoter and the Novel Hotspot Ura4-Aim

The M26 mutation in the ade6 gene of Schizosaccharomyces pombe creates a hot spot of meiotic recombination. A single base substitution, the M26 mutation is situated within the open reading frame, near the 5' end. It has previously been shown that the heptanucleotide sequence 5' ATGACGT 3', which includes the M26 mutation, is required for hot spot activity. The 510-bp ade6-delXB deletion encompasses the promoter and the first 23 bp of the open reading frame, ending 112 bp upstream of M26. Deletion of the promoter in cis to M26 abolishes hot spot activity, while deletion in trans to M26 has no effect. Homozygous deletion of the promoter also eliminates M26 hot spot activity, indicating that the heterology created through deletion of the promoter per se is not responsible for the loss of hot spot activity. Thus, DNA sequences other than the heptanucleotide 5' ATGACGT 3', which must be located at the 5' end of the ade6 gene, appear to be required for hot spot activity. While the M26 hotspot stimulates crossovers associated with M26 conversion, it does not affect the crossover frequency in the intervals adjacent to ade6. The flanking marker ura4-aim, a heterology created by insertion of the ura4(+) gene upstream of ade6, turned out to be a hot spot itself. It shows disparity of conversion with preferential loss of the insertion. The frequency of conversion at ura4-aim is reduced when the M26 hot spot is active 15 kb away, indicating competition for recombination factors by hot spots in close proximity.     

p110α Hot Spot Mutations E545K and H1047R Exert Metabolic Reprogramming Independently of p110α Kinase Activity

The phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) catalytic subunit p110α is the most frequently mutated kinase in human cancer, and the hot spot mutations E542K, E545K, and H1047R are the most common mutations in p110α. Very little is known about the metabolic consequences of the hot spot mutations of p110α in vivo. In this study, we used adenoviral gene transfer in mice to investigate the effects of the E545K and H1047R mutations on hepatic and whole-body glucose metabolism. We show that hepatic expression of these hot spot mutations results in rapid hepatic steatosis, paradoxically accompanied by increased glucose tolerance, and marked glycogen accumulation. In contrast, wild-type p110α expression does not lead to hepatic accumulation of lipids or glycogen despite similar degrees of upregulated glycolysis and expression of lipogenic genes. The reprogrammed metabolism of the E545K and H1047R p110α mutants was surprisingly not dependent on altered p110α lipid kinase activity.     

Identification of a New Genomic Hot Spot of Evolutionary Diversification of Protein Function

Establishment of phylogenetic relationships remains a challenging task because it is based on computational analysis of genomic hot spots that display species-specific sequence variations. Here, we identify a species-specific thymine-to-guanine sequence variation in the Glrb gene which gives rise to species-specific splice donor sites in the Glrb genes of mouse and bushbaby. The resulting splice insert in the receptor for the inhibitory neurotransmitter glycine (GlyR) conveys synaptic receptor clustering and specific association with a particular synaptic plasticity-related splice variant of the postsynaptic scaffold protein gephyrin. This study identifies a new genomic hot spot which contributes to phylogenetic diversification of protein function and advances our understanding of phylogenetic relationships.     

Gene Action of Dollar Spot Resistance in Creeping Bentgrass

The dollar spot disease, incited by Sclerotinia homoeocarpa F.T. Bennet, is one of the most important diseases of creeping bentgrass (Agrostis stolonifera L.) on golf courses. An understanding of the inheritance of dollar spot resistance could enhance genetic improvement efforts in creeping bentgrass. The objectives of this study were to evaluate the response of two creeping bentgrass crosses to two different isolates of S. homoeocarpa, determine gene action and identify number of loci involved in resistance to individual fungal isolates. Parental clones, pseudo F₂, pseudo F₃, BC₁ and BC₂ progenies from two crosses were established in a field trial in a randomized complete block split‐plot design in the fall of 2002. Progeny of each generation (subplots) were inoculated with each of two isolates of S. homoeocarpa (main plots) applied at a rate of 0.25 g/m² of prepared inoculum and evaluated for dollar spot disease. Minimum loci calculations averaged 1.0–2.6. Midparent heterosis calculations were not significant. Backcross population means were closest to the recurrent parent. Generation mean analysis supports a simple additive‐dominance model for both crosses and both isolates, although there was also some evidence of epistatic gene action depending on the cross and the isolate. These results confirm previous research that dollar spot disease is quantitatively inherited and indicate that there may be a few genes interacting in a mainly additive fashion to confer dollar spot disease resistance in creeping bentgrass.     

Using Laser Duty Cycles for Modifying the Performance of Au-NPs/Si Nano Column Hot Spot SERS Sensors

In the current study, the Si nano column layer via pulsed photo chemical etching with different laser pulse duty cycle 30 and 60% using short laser wavelength (405 nm) and laser intensity (100 mW/cm²) was formed and studied. Two types of Si nano column-based plasmonic Au-NP hot spot layers were synthesized and examined successfully as an efficient SERS layer for the detection of the ultra-low concentration of amoxicillin. Si nano columns exposed a great effect on the performance of the Au-NP hot spot SERS sensor showing a strong dependence on the density of the hot spot gaps within the sensitive layer. Enhancement factor (EF) of the Raman signal improved considerably with increasing the density of the hot spot gaps due to the coupling efficiency among the plasmonic Au-NPs and the molecules of amoxicillin within the hot spot regions. EF increased by about four orders of magnitude with decreasing the laser duty cycles due to the increase of the integrated plasmonic Au-NPs into the Si nano column layer.

     

Clustering of Mycobacterium avium subsp. paratuberculosis in Rabbits and the Environment: How Hot Is a Hot Spot?

Clustering of pathogens in the environment leads to hot spots of diseases at local, regional, national, and international levels. Scotland contains regional hot spots of Johne's disease (caused by Mycobacterium avium subsp. paratuberculosis) in rabbits, and there is increasing evidence of a link between paratuberculosis infections in rabbits and cattle. The spatial and temporal dynamics of paratuberculosis in rabbits within a hot spot region were studied with the overall aim of determining environmental patterns of infection and thus the risk of interspecies transmission to livestock. The specific aims were to determine if prevalence of paratuberculosis in rabbits varies temporally between seasons and whether the heterogeneous spatial environmental distribution of M. avium subsp. paratuberculosis on a large scale (i.e., regional hot spots) is replicated at finer resolutions within a hot spot. The overall prevalence of M. avium subsp. paratuberculosis in rabbits was 39.7%; the temporal distribution of infection in rabbits followed a cyclical pattern, with a peak in spring of 55.4% and a low in summer of 19.4%. Spatially, M. avium subsp. paratuberculosis-infected rabbits and, thus, the risk of interspecies transmission were highly clustered in the environment. However, this is mostly due to the clustered distribution of rabbits. The patterns of M. avium subsp. paratuberculosis infection in rabbits are discussed in relation to the host's socioecology and risk to livestock.     

Transposition of mobile dispersed genes (MDG) after substitution of various chromosome pairs in inbred Drosophila melanogaster strains

Transpositions of MDG-1, MDG-3 and copia were detected as a result of crosses of the inbred maladaptive LA stock with laboratory stocks, in order to construct the genomes carrying different combinations of the LA or non-La chromosomal pairs. Changes of the mobile gene distributions were revealed in chromosomes of hybrid genotypes, as compared to parental chromosomal pairs. A trivial source of variability of chromosomal molecular structure ensured by crossing over was excluded by inversions which serve as suppressors of crossing over in corresponding crosses. Multiple transpositions of mobile genes in definite chromosomal sites were detected in genotypes carrying chromosomal pair 2 originated from the LA stock. No such transpositions were observed, when the pair 2 was substituted by the chromosome 2 originated from the Swedish-b line or in control crosses, where the LA stock was not involved. Both LA chromosomes 2 and 3 were shown to be the targets of transpositions. Comparison of hot spot transposition sites of MDG-1, as a result of crosses, with the earlier described rare events of spontaneous transpositions in the LA stock, coupled with its fitness increase, revealed that the hot spot sites were shared in both series of experiments. The data obtained show that transpositions of mobile genetic elements may change the genetic and molecular structure of the chromosome involved in crosses, in spite of suppression of crossing over by inversions usually suggested as a tool for keeping chromosomal genetic structure intact.     

An Implanted Recombination Hot Spot Stimulates Recombination and Enhances Sister Chromatid Cohesion of Heterologous Yacs during Yeast Meiosis

Heterologous yeast artificial chromosomes (YACs) do not recombine with each other and missegregate in 25% of meiosis I events. Recombination hot spots in the yeast Saccharomyces cerevisiae have previously been shown to be associated with sites of meiosis-induced double-strand breaks (DSBs). A 6-kb fragment containing a recombination hot spot/DSB site was implanted onto two heterologous human DNA YACs and was shown to cause the YACs to undergo meiotic recombination in 5-8% of tetrads. Reciprocal exchanges initiated and resolved within the 6-kb insert. Presence of the insert had no detectable effect on meiosis I nondisjunction. Surprisingly, the recombination hot spots acted in cis to significantly reduce precocious sister-chromatid segregation. This novel observation suggests that DSBs are instrumental in maintaining cohesion between sister chromatids in meiosis I. We propose that this previously unknown function of DSBs is mediated by the stimulation of sister-chromatid exchange and/or its intermediates.     

Preferential Binding of Hot Spot Mutant p53 Proteins to Supercoiled DNA In Vitro and in Cells

Hot spot mutant p53 (mutp53) proteins exert oncogenic gain-of-function activities. Binding of mutp53 to DNA is assumed to be involved in mutp53-mediated repression or activation of several mutp53 target genes. To investigate the importance of DNA topology on mutp53-DNA recognition in vitro and in cells, we analyzed the interaction of seven hot spot mutp53 proteins with topologically different DNA substrates (supercoiled, linear and relaxed) containing and/or lacking mutp53 binding sites (mutp53BS) using a variety of electrophoresis and immunoprecipitation based techniques. All seven hot spot mutp53 proteins (R175H, G245S, R248W, R249S, R273C, R273H and R282W) were found to have retained the ability of wild-type p53 to preferentially bind circular DNA at native negative superhelix density, while linear or relaxed circular DNA was a poor substrate. The preference of mutp53 proteins for supercoiled DNA (supercoil-selective binding) was further substantiated by competition experiments with linear DNA or relaxed DNA in vitro and ex vivo. Using chromatin immunoprecipitation, the preferential binding of mutp53 to a sc mutp53BS was detected also in cells. Furthermore, we have shown by luciferase reporter assay that the DNA topology influences p53 regulation of BAX and MSP/MST1 promoters. Possible modes of mutp53 binding to topologically constrained DNA substrates and their biological consequences are discussed.     

Hot spot of recombination within DXS164 in the Duchenne muscular dystrophy gene.

The DMD gene, which spans more than 2,000 kbp, has been assigned to band Xp21 of the X chromosome. Two subclones (PERT 87-1 and PERT 87-15) of the intragenic locus DXS164 physically are separated by approximately 60 kbp. Linkage studies were done in 49 informative DMD families by using the LINKAGE program. Crossing-over between the loci studied occurred in four families. A recombination rate of 4% (support interval [Zmax-1] 1%-10%), which was 54 (support interval 14-135-fold) times higher than expected, was found with a maximum lod score of 13.50. These data suggest a hot spot for recombination within DXS164.     

Molecular Analysis of the Hot Spot Region Related to Length Mutations in Wheat Chloroplast Dnas. I. Nucleotide Divergence of Genes and Intergenic Spacer Regions Located in the Hot Spot Region

The nucleotide divergence of chloroplast DNAs around the hot spot region related to length mutation in Triticum (wheat) and Aegilops was analyzed. DNA sequences (ca. 4.5 kbp) of three chloroplast genome types of wheat complex were compared with one another and with the corresponding region of other grasses. The sequences region contained rbcL and psaI, two open reading frames, and a pseudogene, rpl23' (pseudogene for ribosomal protein L23) disrupted by AT-rich intergic spacer regions. The evolution of these genes in the closely related wheat complex is characterized by nonbiased nucleotide substitutions in terms of being synonymous/nonsynonymous, having A-T pressure transitions over transversions, and frequent changes at the third codon position, in contrast with the gene evolution among more distant plant groups where biased nucleotide substitutions have frequently occurred. The sequences of these genes had diverged almost in proportion to taxonomic distance. The sequence of the pseudogene rpl23' changed approximately two times faster than that of the coding region. Sequence comparison between the pseudogene and its protein-coding counterpart revealed different degrees of nucleotide homology in wheat, rice and maize, suggesting that the transposition timing of the pseudogene differed and/or that different rates of gene conversion operated on the pseudogene in the cpDNA of the three plant groups in Gramineae. The intergenic spacer regions diverged approximately ten times faster than the genes. The divergence of wheat from barley, and that from rice are estimated based on the nucleotide similarity to be 1.5, 10 and 40 million years, respectively.     

Localized breakdown in linkage disequilibrium does not always predict sperm crossover hot spots in the human MHC class II region

To investigate the relationship between meiotic crossover hot spots and block-like linkage disequilibrium (LD), we have extended our high-resolution studies of the human MHC class II region to a 90-kb segment upstream of the HLA-DOA gene. LD blocks in this region are not as well defined as in the neighboring 210-kb DNA segment but do show two regions of LD breakdown in which coalescent analysis indicates substantial historical recombination. Sperm crossover analysis of one region revealed a novel localized hot spot similar in intensity and morphology to most other MHC hot spots. Crossovers at this hot spot are not obviously affected by a large insertion/deletion polymorphism near the hot spot. The second region of LD breakdown, within the DPB1 gene, shows an extremely low level of sperm crossover activity and does not contain a sperm crossover hot spot. These results highlight the complexity of LD patterns and the importance of experimentally verifying crossover hot spots.     

Escherichia coli RecBC pseudorevertants lacking chi recombinational hotspot activity

Pseudorevertants of an Escherichia coli exonuclease V (RecBC enzyme)-negative mutant have been isolated after ethyl methane sulfonate mutagenesis of a recC73 (presumed missense) mutant. The remedial mutations in each of the four pseudorevertants studied in detail map and complement as recC mutations. By several criteria, such as recombination proficiency, support of phage growth, RecBC nuclease activity, and cell viability, the pseudorevertants appear to have regained partially or completely various aspects of RecBC activity. However, chi recombinational hotspots, which stimulate exclusively the RecBC pathway of recombination, have no detectable activity in lambda vegetative crosses in the pseudorevertants. The properties of these mutants, in which the RecBC pathway of recombination is active yet in which chi is not active, are consistent with the hypothesis that wild-type RecBC enzyme directly interacts with chi sites; alternatively, the mutants may block or bypass the productive interaction of another recombinational enzyme with chi.     

Hot Spots, Cold Spots, and the Geographic Mosaic Theory of Coevolution

Species interactions commonly coevolve as complex geographic mosaics of populations shaped by differences in local selection and gene flow. We use a haploid matching-alleles model for coevolution to evaluate how a pair of species coevolves when fitness interactions are reciprocal in some locations ("hot spots") but not in others ("cold spots"). Our analyses consider mutualistic and antagonistic interspecific interactions and a variety of gene flow patterns between hot and cold spots. We found that hot and cold spots together with gene flow influence coevolutionary dynamics in four important ways. First, hot spots need not be ubiquitous to have a global influence on evolution, although rare hot spots will not have a disproportionate impact unless selection is relatively strong there. Second, asymmetries in gene flow can influence local adaptation, sometimes creating stable equilibria at which species experience minimal fitness in hot spots and maximal fitness in cold spots, or vice versa. Third, asymmetries in gene flow are no more important than asymmetries in population regulation for determining the maintenance of local polymorphisms through coevolution. Fourth, intraspecific allele frequency differences among hot and cold spot populations evolve under some, but not all, conditions. That is, selection mosaics are indeed capable of producing spatially variable coevolutionary outcomes across the landscapes over which species interact. Altogether, our analyses indicate that coevolutionary trajectories can be strongly shaped by the geographic distribution of coevolutionary hot and cold spots, and by the pattern of gene flow among populations.     

珊瑚礁生态系统模型研究现状及热点分析

以Web of Science数据库中的Web of Science ~(TM)核心合集和中国期刊全文数据库为数据源,对2018年10月前国内外发表的珊瑚礁生态系统模型研究相关文献进行计量分析,评估该领域的研究现状及热点。研究表明:国外珊瑚礁生态系统模型研究起始于1974年,相关文献的发文量整体呈大幅度增长趋势;国内研究于1997年起步,发文量增长较缓慢,近三年显著增加。高频关键词统计分析可以看出,国外对于珊瑚礁中生活的各种生物及其生态分布研究较为广泛,生态系统的生物群落多样性是该领域的研究热点,关于珊瑚礁生态系统风险、威胁、胁迫和修复的模型研究近几年有所起色,在今后一段时间内将是该领域的主要研究趋势。国内相关研究局限于珊瑚礁地质环境与波浪传播等方面,应在扩大研究范围的同时,将重点放在珊瑚礁生态系统退化诊断与修复的探究上。     

三七总皂苷的药用生物学活性研究进展

三七总皂苷是中药三七的主要有效成分,对中枢神经系统、心血管系统、血液系统等具有抗炎、抗纤维化、抗衰老、抗肿瘤等方面的生物学活性,是现代中药、药理等领域的研究热点.近年来随着研究的不断深入,三七总皂苷的生物学活性被进一步发现,其用途也越来越广.本文就三七总皂苷在生物学方面的研究情况和用途进行综述,并对其前景予以讨论.