A three-dimensional model of the U1 small nuclear ribonucleoprotein particle

Most of the pre-mRNAs in the eukaryotic cell are comprised of protein-coding exons and non-protein-coding introns. The introns are removed and the exons are ligated together, or spliced, by a large, macromolecular complex known as the spliceosome. This RNA-protein assembly is made up of five uridine-rich small nuclear RNAs (U1-, U2-, U4-, U5- and U6-snRNA) as well over 300 proteins, which form small nuclear ribonucleoprotein particles (snRNPs). Initial recognition of the 5′ exon/intron splice site is mediated by the U1 snRNP, which is composed of the U1 snRNA as well as at least ten proteins. By combining structural informatics tools with the available biochemical and crystallographic data, we attempted to simulate a complete, three dimensional U1 snRNP from the silk moth, Bombyx mori. Comparison of our model with empirically derived crystal structures and electron micrographs pinpoints both the strengths and weaknesses in the in silico determination of macromolecular complexes. One of the most striking differences between our model and experimentally generated structures is in the positioning of the U1 snRNA stem-loops. This highlights the continuing difficulties in generating reliable, complex RNA structures; however, three-dimensional modeling of individual protein subunits by threading provided models of biological significance and the use of both automated and manual docking strategies generated a complex that closely reflects the assembly found in nature. Yet, without utilizing experimentally-derived contacts to select the most likely docking scenario, ab initio docking would fall short of providing a reliable model. Our work shows that the combination of experimental data with structural informatics tools can result in generation of near-native macromolecular complexes.     

Functional analysis of a miRNA‐like small RNA derived from Bombyx mori cytoplasmic polyhedrosis virus

Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is a major pathogen of the economic insect silkworm, Bombyx mori. Virus‐encoded microRNAs (miRNAs) have been proven to play important roles in host–pathogen interactions. In this study we identified a BmCPV‐derived miRNA‐like 21 nt small RNA, BmCPV‐miR‐1, from the small RNA deep sequencing of BmCPV‐infected silkworm larvae by stem‐loop quantitative real‐time PCR (qPCR) and investigated its functions with qPCR and lentiviral expression systems. Bombyx mori inhibitor of apoptosis protein (BmIAP) gene was predicted by both target prediction software miRanda and Targetscan to be one of its target genes with a binding site for BmCPV‐miR‐1 at the 5′ untranslated region. It was found that the expression of BmCPV‐miR‐1 and its target gene BmIAP were both up‐regulated in BmCPV‐infected larvae. At the same time, it was confirmed that BmCPV‐miR‐1 could up‐regulate the expression of BmIAP gene in HEK293T cells with lentiviral expression systems and in BmN cells by transfecting mimics. Furthermore, BmCPV‐miR‐1 mimics could up‐regulate the expression level of BmIAP gene in midgut and fat body in the silkworm. In the midgut of BmCPV‐infected larvae, BmCPV‐miR‐1 mimics could be further up‐regulated and inhibitors could lower the virus‐mediated expression of BmIAP gene. With the viral genomic RNA segments S1 and S10 as indicators, BmCPV‐miR‐1 mimics could up‐regulate and inhibitors down‐regulate their replication in the infected silkworm. These results implied that BmCPV‐miR‐1 could inhibit cell apoptosis in the infected silkworm through up‐regulating BmIAP expression, providing the virus with a better cell circumstance for its replication.     

激光诱导家蚕孤雌生殖的研究 Ⅳ.Lamp 1的遗传学研究

通过杂交和解剖试验,我们发现Ｌａｍｐ１为一常染色体（Ⅱ）和Ｗ易位片段同时缺失或突变的斑纹限性孤雌生殖突变体。与亲本回交后代分离比,１浓普♀：３正常普♀或：３淡普♀或：１素蚕♀或１＋ｐ＋ｐ／：２Ｗ＋ｐ／：１＋ｐ＋ｐ／ＺＺ：１ＰＰ／：ＺＷ＋ｐ／ＺＺ：１ＰＰ／ＺＺ。联会复合体分析仅浓普斑雌蚕表型个体中发现染色体易位片段。     

BmCaspase-8-like regulates autophagy by suppressing BmDREDD-mediated cleavage of BmATG6

Autophagy plays an important role in tissue remodeling during insect development. The interplay between autophagy-related (ATG) proteins and caspases regulates the autophagic activity of ATGs, thereby modulating the process of autophagy. Our previous study characterized BmCaspase-8-like (BmCasp8L) as a caspase suppressor that inhibits apoptosis and immune signaling by suppressing the activation of death-related ced-3/Nedd2-like caspase (DREDD), a caspase-8 homolog in silkworm. In this study, we explored the regulatory role of BmCasp8L in autophagy. We found that the expression of Bmcasp8l increased from the late spinning stage to the pupa stage in the posterior silk gland (PSG), correlating with the expression patterns of Bmatg8 and Bmatg6. RNA interference-mediated downregulation of BmCasp8L expression significantly decreased starvation-induced autophagic influx as determined by the levels of BmATG8–phosphatidylethanolamine and the percentage of cells displaying punctate enhanced green fluorescent protein-BmATG8. Conversely, the overexpression of BmCasp8L significantly increased autophagic influx. We also found that BmCasp8L underwent autophagic degradation induced by starvation and that it was colocalized with BmATG8. Lastly, we demonstrated that BmDREDD attenuated autophagy and BmCasp8L suppressed BmDREDD-mediated cleavage of BmATG6. Taken together, our results demonstrated that BmCasp8L is a novel proautophagic molecule which suppresses BmDREDD-mediated cleavage of BmATG6 and is a target for autophagy.     

Fumarate mitigates disruption induced by fenpropathrin in the silkworm Bombyx mori (Lepidoptera): A metabolomics study

The silkworm Bombyx mori L. is a model organism of the order Lepidoptera. Understanding the mechanism of pesticide resistance in silkworms is valuable for Lepidopteran pest control. In this study, comparative metabolomics was used to analyze the metabolites of 2 silkworm strains with different pesticide resistance levels at 6, 12, and 24 h after feeding with fenpropathrin. Twenty-six of 27 metabolites showed significant differences after fenpropathrin treatment and were classified into 6 metabolic pathways: glycerophospholipid metabolism, sulfur metabolism, glycolysis, amino acid metabolism, the urea cycle, and the tricarboxylic acid (TCA) cycle. After analyzing the percentage changes in the metabolic pathways at the 3 time points, sulfur metabolism, glycolysis, and the TCA cycle showed significant responses to fenpropathrin. Confirmatory experiments were performed by feeding silkworms with key metabolites of the 3 pathways. The combination of iron(II) fumarate + folic acid (IF-FA) enhanced fenpropathrin resistance in silkworms 6.38 fold, indicating that the TCA cycle is the core pathway associated with resistance. Furthermore, the disruption of several energy-related metabolic pathways caused by fenpropathrin was shown to be recovered by IF-FA in vitro. Therefore, IF-FA may have a role in boosting silkworm pesticide resistance by modulating the equilibrium between the TCA cycle and its related metabolic pathways.     

DECISION TOOLS FOR MULBERRY THRIPS PSEUDODENDROTHRIPS MORI (NIWA, 1908) MANAGEMENT IN SERICULTURAL REGIONS; AN OVERVIEW

Abstract Mulberry thrips Pseudodendrothrips mori (Niwa, 1908) is a major pest of mulberry trees recorded from different sericultural regions of the world. The thrips infestation affects the qualitative and quantitative characters of mulberry leaf, by direct feeding damage to leaves and the ingestion of sap, which in turn affects the silkworm cocoon crop. This is most harmful in dry climates and seasons when heavily attacked plants lose moisture heavily. Under these conditions infestation can seriously deplete yields. The seasonal population fluctuation and the degree of damage caused to the host plant are influenced by various environmental factors including climate, host-plant variety, topography, soil type, and management regimes. This article attempts to review all available documents on mulberry thrips and to discuss the practical approaches for best control of this pest.     

食管癌病人运铁蛋白(Tf)和组特异性成份(Gc)亚型分布的研究

应用超薄层聚丙烯酰胺凝胶等电聚焦电泳法, 分析了潮汕地区216例无血缘关系、临床上诊断为食管癌的病人和216例健康人的运铁蛋白(Tf)亚型分布情况, 结果发现:食管癌病人组TfC1C1纯合子频率为0.2639,Tf*Cl基因频率为0.4745,显著低于正常人组(分别为0.4352和0.6227,均为P<0.0 01);同时,食管癌病人组TfC2C2纯合子频率为0.2278,Tf*C2基因频率为0. 4977,显著高于正常对照组(分别为0.1852,P<0.05,和0.3634,P<0.001)。应用超薄层聚丙烯酰胺凝胶等电聚焦结合免疫固定法,分析了潮汕地区21 7例无血缘关系的临床上诊断为食管癌病人和 217例健康人的组特异性成份(Gc)亚型的分布,发现两组间无显著性差异。     

Developmental profiles of wing imaginal discs of flügellos(fl), a wingless mutant of the silkworm, Bombyx mori

 Mutations at the flügellos (fl) locus in Bombyx mori give rise to wingless pupae and moths. To understand the developmental steps responsible for the fl wing defect, we compared the morphological changes and protein synthesis profiles between fl and wild-type (WT) wing discs during larval development. Morphologically, the four wing discs in the fl homozygote larva developed normally at least until the fourth instar, but they were slightly smaller than those of the WT. After the last larval ecdysis, wing epithelial invagination and tracheal migration into the lacunar spaces evidently occurred in the WT wing discs. However, there was no apparent morphological change in fl discs through the fifth instar. The fl wing discs cultured in medium containing 20-hydroxyecdysone (20E) did not grow and develop, although the WT wing discs extended and differentiated under the same conditions. A comparison of protein synthesis in the wing discs revealed that several bands were differentially expressed between the fl and WT. A 41-kDa band expressed abundantly from larval to pharate pupal stages in the WT wing discs was rarely observed in fl discs. Furthermore, in vitro culture studies showed that the 41-kDa protein was induced by 20E and specifically synthesized in WT wing discs after the wandering stage, but not in fl discs. The wing-specific protein synthesis and morphogenesis in fl wing discs may be blocked due to aberrant expression of the fl gene. Received: 6 November 1996 / Accepted: 5 February 1997