Functional Inactivation of Drosophila GCK Orthologs Causes Genomic Instability and Oxidative Stress in a Fly Model of MODY-2

Maturity-onset diabetes of the young (MODY) type 2 is caused by heterozygous inactivating mutations in the gene encoding glucokinase (GCK), a pivotal enzyme for glucose homeostasis. In the pancreas GCK regulates insulin secretion, while in the liver it promotes glucose utilization and storage. We showed that silencing the Drosophila GCK orthologs Hex-A and Hex-C results in a MODY-2-like hyperglycemia. Targeted knock-down revealed that Hex-A is expressed in insulin producing cells (IPCs) whereas Hex-C is specifically expressed in the fat body. We showed that Hex-A is essential for insulin secretion and it is required for Hex-C expression. Reduced levels of either Hex-A or Hex-C resulted in chromosome aberrations (CABs), together with an increased production of advanced glycation end-products (AGEs) and reactive oxygen species (ROS). This result suggests that CABs, in GCK depleted cells, are likely due to hyperglycemia, which produces oxidative stress through AGE metabolism. In agreement with this hypothesis, treating GCK-depleted larvae with the antioxidant vitamin B6 rescued CABs, whereas the treatment with a B6 inhibitor enhanced genomic instability. Although MODY-2 rarely produces complications, our data revealed the possibility that MODY-2 impacts genome integrity.     

XIII. Yeast sequencing reports. Cloning and sequencing of the NES24 gene of Saccharomyces cerevisiae

We have cloned NES24 using a temperature-sensitive nes24-1 mutant as a host and sequenced a 3162 bp XhoI-EcoRI DNA fragment containing the NES24 gene. Computer analysis revealed that this segment contains a 1806 bp open reading frame which is needed for complementation of the nes24-1 mutation. We found SUP8 in the region upstream of the NES24 gene, placing the NES24 gene on chromosome XIII. A protein homology search indicated that NES24 encodes a new protein. The disruption of the NES24 gene resulted in temperature-sensitive growth. The sequence has been deposited in DDBJ/EmBL/GenBank data bases under Accession Number D15052.     

XV. Yeast sequencing reports. Isolation and DNA sequence of the STE13 gene encoding dipeptidyl aminopeptidase

We have isolated a mutant which exhibits partial constitutivity for a -specific gene expression in α cells. The wild-type gene was cloned and demonstrated to be allelic to the STE13 gene, which encodes the dipeptidyl aminopeptidase involved in processing of the α-factor prepropheromone. Thus, the mating defect of the ste13 mutations in α cells may result both from the production of incompletely processed α-factor and from the increased expression of a -specific genes. The STE13 open reading frame of 931 amino acids contains a putative membrane-spanning segment near its amino terminus and is 31% identical to a second yeast dipeptidyl aminopeptidase (DAP2). A null mutant of STE13 has been constructed: it is viable and sporulation-proficient. The sequence has been deposited in the GenBank data library under Accession Number L21944.     

II. Yeast sequencing reports. Sequence of a segment of yeast chromosome II shows two novel genes,one almost entirely hydrophobic and the other extremely asparagine-serine rich

A 3·2 kb EcoRI fragment of yeast Saccharomyces cerevisiae was entirely sequenced. Two new open reading frames were identified. The first is extremely hydrophobic, and would likely be an integral membrane protein. It has significant similarity to only one reported gene, a gene of unknown function from Drosophila melanogaster. The second ORF is asparagine-rich and very serine-rich, with a remarkable stretch of nearly 26 consecutive asparagine residues comprised of the same codon. It has no significant similarity to any reported gene. The fragment maps to chromosome II on the left arm between the CDC27 and ILS1 loci. The nucleotide sequence reported in this paper has been deposited in the GenBank database with the Accession Number M89908.     

The complete sequence of a 10.8 kb segment distal of SUF2 on the right arm of chromosome III from Saccharomyces cerevisiae reveals seven open reading frames including the RVS161, ADP1 and PGK genes.

We have entirely sequenced a 10,835 bp segment of the right arm from chromosome III contained in the J11D and J11D-K3B GF clones. The segment contains seven open reading frames longer then 100 amino acids. Three of them, RVS161 (Urdaci et al., 1990; Crouzet et al., 1991), ADP1 (Purnelle et al., 1991) and PGK1 (Hitzeman et al., 1982) have been described previously. YCR10C encodes a putative membrane protein. YCR8W (encoding a putative protein kinase) and YCR14C extend inside the D10H (Skala et al., 1991) and 62B5-2D clones respectively. Four ARS elements previously reported by Palzkill et al. (1986) are located between RVS161 and YCR10C.     

Analysis of the MSS51 region on chromosome XII of Saccharomyces cerevisiae.

We have localized gene MSS51 on chromosome XII of Saccharomyces cerevisiae between the RDN1 and CDC42 loci. 'Head to head' with MSS51 is another gene, QRI5, the function of which is unknown. However, the proximity of these genes, the structure of the intergenic region and the presence of an ABF1 binding site right in the middle of this region suggest that the MSS51 and QRI5 expressions are submitted to a common regulatory process.     

The three-dimensional architecture of the mitotic spindle,analyzed by confocal fluorescence and electron microscopy

Fluorescence microscopy techniques have become important tools in mitosis research. The well-known disadvantages of fluorescence microscopy, rapid bleaching, phototoxicity and out-of-focus contributions blurring the in-focus image are obstacles which still need to be overcome. Confocal fluorescence microscopy has the potential to improve our capabilities of analyzing cells, because of its excellent depth-discrimination and image processing power. We have been using a confocal fluorescence microscope for the study of the mechanism of poleward chromosome movement, and report here (1) a cell preparation technique, which allows labeling of fixation sensitive spindle antigens with acceptable microtubule preservation; (2) the use of image processing methods to represent the spatial distribution of various labeled elements in pseudocolour; (3) a novel immunoelectron microscopic labeling method for microtubules, which allows the visualization of their distribution in semithin sections at low magnification; and (4) a first attempt to study microtubule dynamics with a confocal fluorescence microscope in living cells, microinjected with rhodamine labeled tubulin. Our experience indicates that confocal fluorescence microscopy provides real advantages for the study of spatial colocalization of antigens in the mitotic spindle. It does not, however, overcome the basic limits of resolution of the light microscope. Therefore, it has been necessary to use an electron microscopic method. Our preliminary results with living cells show that it is possible to visualize the entire microtubule network in stereo, but that the sensitivity of the instrument is still too low to perform dynamic time studies. It will be worthwhile to further develop this new type of optical instrumentation and explore its usefulness on both fixed and living cells.     

野生黑腹果蝇引种培养与唾腺染色体观察

对佳木斯地区的野生黑腹果蝇（Drosophila melanogaster）进行采集,并在实验室内进行培养训化,用常规果蝇培养基进行培养．选用卡宝品红染色法进行唾腺染色体临时装片的制备,观察其染色效果,与常规方法进行比较,效果比较好,并应用于遗传学教学．     

交互更新模式的量子遗传算法的几何约束求解

目的针对传统量子遗传算法无法充分利用种群中未成熟个体信息的不足,提出了基于交互更新模式的量子遗传算法(IUMQGA)并应用于几何约束求解中。方法几何约束问题的约束方程组可转化为优化模型,因此约束求解问题可以转化为优化问题。采用将遗传算法与量子理论相结合的量子遗传算法,使用双串量子染色体结构,使用交互更新策略将遗传算法中的交叉操作利用量子门变换来实现,根据不同情况采用不同的交互更新策略。这里的交互,指的是两个个体进行信息交换的过程,该过程用以产生新的个体。这不仅增加了个体间信息的交换而且充分利用了种群中未成熟个体的信息,提高了算法的收敛速度。结果通过非线性方程实例和几何约束实例测试并与其他方法比较表明,基于交互更新模式的量子遗传算法求解几何约束问题具有更好的求解精度和求解速率。双圆外公切线问题实例中,IUMQGA算法比QGA算法稳定;单圆填充问题和双圆外公切线问题实例中,通过实验求得各变量的最优值与其相应的精确值的误差在1E-2以下。结论采用交互更新模式的量子遗传算法可以很好地求解几何约束问题。     

交互式遗传算法在作曲中的应用

对交互式遗传算法作曲进行了一定的探讨.介绍了用遗传算法进行作曲的知识表示.通过将给定的乐谱转化成相应的编码,采用交互式遗传算法中的选择,交叉和变异算子产生新的乐曲.对交互式遗传算法作曲进行了仿真实验,讨论了实验步骤,给出了实验结果.仿真结果表明,设计的作曲系统可以产生出令人感兴趣的乐曲,具有一定的实际意义.