茄科作物青枯菌NADH脱氢酶F亚基在细胞运动和致病性中的功能研究

[目的]劳尔氏菌（Ralstonia solanacearum）在茄科作物上引起严重的细菌性青枯病,本研究旨在发掘青枯劳尔氏菌与致病相关的基因。[方法]利用Tn5转座子构建随机插入突变体,分析生物膜形成、细胞运动和致病性;对有表型变化的突变体,运用TAIL-PCR方法鉴定Tn5插入位点,确定所突变的基因。[结果]以模式菌株GMI000为出发菌,总共获得了400个突变体,其中2个突变体不能形成生物膜,在软琼脂平板上的运动能力下降;接种感病番茄植物,这2个突变体都不能引起萎焉症状。TAIL-PCR结果显示,2个突变体的Tn5插入位点都在NADH脱氢酶F亚基（nuoF）中,距离翻译起始位点分别为103-bp和225-bp。ripAY基因启动子推动的nuoF基因互补载体,完全恢复了2个突变体的表型。[结论]NADH脱氢酶复合物是微生物呼吸电子传递链中的第一步催化酶。我们的结果表明,NADH脱氢酶复合物对R.solanacearum生物膜形成、细胞运动和致病性也有重要作用。     

Blocking P2X7 receptor ameliorates oxidized LDL-mediated podocyte apoptosis

Molecular Biology Reports - The purpose of our research is to elucidate whether oxLDL activates P2X7R in cultured human podocytes and if the activation of P2X7R leads to podocyte apoptosis....     

Overexpression of the lncRNA FER1L4 inhibits paclitaxel tolerance of ovarian cancer cells via the regulation of the MAPK signaling pathway

To determine how the lncRNA FER1L4 in ovarian cancer cells influences paclitaxel (PTX) resistance, we examined the expression level of FER1L4 in human ovarian epithelial cell lines IOSE80 and HOSEpiC and human ovarian cancer cell lines OVCAR-3, Caov-3, and SKOV3 through RNA isolation and quantitative polymerase chain reaction (qRT-PCR). SKOV3 cell lines were treated with PTX. The cell survival rate and apoptosis rate of SKOV3 and SKOV3-PR at different PTX dose levels were evaluated. Next, qRT-PCR was performed to detect the expression of FER1L4 in SKOV3 and SKOV3-PR cell lines. SKOV3-PR cell lines were transfected with pcDNA3.1 as the control group (SKOV3-PR/pcDNA3.1) or pcDNA3.1-FER1L4 to upregulate the expression level of FER1L4 (SKOV3-PR/pcDNA3.1-FER1L4). The level of cell survival, apoptosis, and colony formation were compared between the two groups using MTT, flow cytometry analysis, and colony formation assay. To reveal the molecular mechanism, we measured the relative protein phosphorylation level of ERK and MAPK in SKOV3, SKOV3-PR, SKOV3-PR/pcDNA3.1, and SKOV3-PR/pcDNA3.1-FER1L4 groups using an enzyme-linked immunosorbent assay. The effects of SB203580 (a p38 MAPK inhibitor) on PTX were also investigated to reveal the function of the MAPK pathway on the PTX tolerance of SKOV3. In comparison with normal ovarian epithelial cells, FER1L4 was downregulated. The FER1L4 level was decreased in human ovarian cancer cells with drug resistance than in common ovarian cancer cells. The upregulation of FER1L4 could promote the PTX sensitivity of ovarian cancer cells. The increased level of FER1L4 could suppress the PTX resistance of ovarian cancer cells through the inhibition of the MAPK signaling pathway.     

拟南芥花粉管与柱头互作的乙醇代谢耦合模型

依据拟南芥公开的代谢途径数据库,构建了基于酶与酶的拟南芥代谢网络模型。利用拟南芥花粉管与柱头互作过程中的转录组数据,挖掘出花粉管与柱头在互作过程中的特异表达基因,进一步将特异表达的酶基因匹配到已构建的拟南芥代谢网络中,根据网络拓扑模型中的节点(酶)之间的共表达关联性,最后给出了一个拟南芥花粉管与柱头互作的乙醇代谢耦合模型。     

Exostosin glycosyltransferase 1 reduces porcine reproductive and respiratory syndrome virus infection through proteasomal degradation of nsp3 and nsp5

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a serious threat to the swine industry worldwide. Exostosin glycosyltransferase 1 (EXT1), an enzyme involved in the biosynthesis of heparin sulfate, has also been reported to be a host factor essential for a wide variety of pathogens. However, the role of EXT1 in PRRSV infection remains uncharted. Here, we identified that PRRSV infection caused an increase of EXT1 expression. EXT1 knockdown promoted virus infection, whereas its overexpression inhibited virus infection, suggesting an inhibitory function of EXT1 to PRRSV infection. We found that EXT1 had no effects on the attachment, internalization, or release of PRRSV but did restrict viral RNA replication. EXT1 was determined to interact with viral nonstructural protein 3 (nsp3) and nsp5 via its N-terminal cytoplasmic tail and to enhance K48-linked polyubiquitination of these two nsps to promote their degradation. Furthermore, the C-terminal glycosyltransferase activity domain of EXT1 was necessary for nsp3 and nsp5 degradation. We also found that EXT2, a EXT1 homolog, interacted with EXT1 and inhibited PRRSV infection. Similarly, EXT1 effectively restricted porcine epidemic diarrhea virus and porcine enteric alphacoronavirus infection in Vero cells. Taken together, this study reveals that EXT1 may serve as a broad-spectrum host restriction factor and suggests a molecular basis for the potential development of therapeutics against PRRSV infection.     

棘冠海星暴发及其对珊瑚礁的生态影响研究进展

棘冠海星的反复暴发是导致印度—太平洋区域珊瑚礁生态系统退化的最主要原因之一。然而,我国对棘冠海星的研究非常有限。本文综述了国内外关于棘冠海星及其暴发的生态影响和应对策略的研究进展,得出以下主要结论：1)雌性棘冠海星个体每年产卵数量高达50万—2亿个,环境因素变化只要导致幼虫和幼体存活率的轻微提高,成体就将得到大量补充;2)棘冠海星暴发的阈值为1000—1500个/km²,暴发周期为10—27 a,每次暴发持续1—10 a,最终可能以“种群集体感染疾病而崩溃”结束;3)棘冠海星暴发对印度洋及太平洋东部和北部珊瑚礁的破坏性非常小,却直接导致太平洋的西部和南部珊瑚礁90%以上的珊瑚死亡,并通过改变珊瑚群落组成、减少珊瑚和鱼类多样性而对珊瑚礁产生间接影响;4)关于棘冠海星暴发原因的假说中“陆地营养物质输入假说”和“捕食者过度捕捞假说”得到了最普遍的认可,但都不能解释所有的暴发事件;5)应对棘冠海星暴发的主要策略有改善水质、设立保护区、投放天敌和人工清理等,其中人工清理是最直接有效的策略,但迄今并没有发现可长期抑制棘冠海星暴发的方法。因此,急需加强对棘冠海星的深入研究,探查...     

CRL4-DCAF8L1 Regulates BRCA1 and BARD1 Protein Stability

BRCA1 is frequently down-regulated in breast cancer, the underlying mechanism is unclear. Here we identified DCAF8L1, an X-linked gene product, as a DDB1-Cullin associated Factor (DCAF) for CUL4 E3 ligases to target BRCA1 and BARD1 for proteasomal degradation. Forced expression of DCAF8L1 caused reduction of BRCA1 and BARD1, and impaired DNA damage repair function, conferring increased sensitivity to irradiation and DNA damaging agents, as well as Olaparib, a PARPi anticancer drug; while depletion of DCAF8L1 restored BRCA1 and suppressed the growth of its xenograft tumors. Furthermore, the expression of DCAF8L1 was induced in human H9 ES cells during transition from primed to naïve state when Xi chromosome was reactivated. Aberrant expression of DCAF8L1 was observed in human breast fibroadenoma and breast cancer. These findings suggest that CRL4^DCAF8L1 is an important E3 ligase that may participate in the development of breast cancer, probably through regulating the stability of BRCA1 and BARD1 tumor suppressor, linking BRCA1 and X chromosome inactivation to breast carcinogenesis.     

抑咽侧体素基因在大猿叶虫生殖滞育准备中的功能分析

[目的]本研究旨在明确大猿叶虫Colaphellus bowringi促咽侧体素(allatotropin,AT)和抑咽侧体素(allatostatin,AST)基因的分子特征,分析其在该虫生殖和滞育过程中的表达差异,并探究其在大猿叶虫生殖滞育准备中的功能.[方法]利用前期建立的大猿叶虫转录组数据库,鉴定大猿叶虫CbA...