Interactive effects of temperature and concentration of the flavonol rutin on growth, molt and food utilization of Manduca sexta caterpillars

A factorial experiment tested the effects of varying concentrations of the flavonol rutin and daytime temperatures of 20 and 30°C on growth, molting and food utilization efficiencies of third instar tobacco hornworms (Manduca sexta (L.)). Cool temperature prolonged both the growth (=feeding) and non-feeding periods and consequently the relative consumption rates (RCR) and relative growth rates (RGR). Temperature had no impact on the amount of food consumed and the utilization indices of efficiency of conversion of ingested food and efficiency of conversion of digested food to larval biomass. But rutin was more concentrated in the frass of the larvae at the warm temperature. With increasing levels of rutin in the diet, the efficiency of conversion of ingested food tended to decline. Rutin reduced RCR and RGR. At the cool temperature, rutin increased the time spent in the first portion of the non-feeding period disproportionately. Analysis of growth rate intervals within the growth period indicated that at the cool temperature rutin had no discernible impact over the first half of the growth period, during which developmental competence to molt is likely achieved. Overall, these results indicate an overlap in the growth and molting phases and suggest that rutin interferes with physiological processes at the time of molt initiation, with these effects magnified at a cool temperature.     

Structure and functions of the translation initiation factor eIF4E and its role in cancer development and treatment

In eukaryotic cells, protein synthesis is a complex and multi-step process that has several mechanisms to start the translation including cap-dependent and cap-independent initiation. The translation control of eukaryotic gene expression occurs principally at the initiation step. In this context, it is critical that the eukaryotic translation initiation factor eIF4E bind to the 7-methylguanosine (m7G) cap present at the 5′-UTRs of most eukaryotic mRNAs. Combined with other initiation factors, eIF4E mediates the mRNA recruitment on ribosomes to start the translation. Moreover, the eIF4E nuclear bodies are involved in the export of specific mRNAs from the nucleus to the cytoplasm. In this review, we focus on the eIF4E structure and its physiological functions, and describe the role of eIF4E in cancer development and progression and the current therapeutic strategies to target eIF4E.     

Translation acrobatics: how cancer cells exploit alternate modes of translational initiation

Recent work has brought to light many different mechanisms of translation initiation that function in cells in parallel to canonical cap‐dependent initiation. This has important implications for cancer. Canonical cap‐dependent translation initiation is inhibited by many stresses such as hypoxia, nutrient limitation, proteotoxic stress, or genotoxic stress. Since cancer cells are often exposed to these stresses, they rely on alternate modes of translation initiation for protein synthesis and cell growth. Cancer mutations are now being identified in components of the translation machinery and in cis‐regulatory elements of mRNAs, which both control translation of cancer‐relevant genes. In this review, we provide an overview on the various modes of non‐canonical translation initiation, such as leaky scanning, translation re‐initiation, ribosome shunting, IRES‐dependent translation, and m⁶A‐dependent translation, and then discuss the influence of stress on these different modes of translation. Finally, we present examples of how these modes of translation are dysregulated in cancer cells, allowing them to grow, to proliferate, and to survive, thereby highlighting the importance of translational control in cancer.     

Role of GhHDA5 in H3K9 deacetylation and fiber initiation in Gossypium hirsutum

Cotton fibers are single‐celled trichomes that initiate from the epidermal cells of the ovules at or before anthesis. Here, we identified that the histone deacetylase (HDAC ) activity is essential for proper cotton fiber initiation. We further identified 15 HDAC s homoeologs in each of the A‐ and D‐subgenomes of Gossypium hirsutum . Few of these HDAC homoeologs expressed preferentially during the early stages of fiber development [?1, 0 and 6 days post‐anthesis (DPA )]. Among them, GhHDA 5 expressed significantly at the time of fiber initiation (?1 and 0 DPA). The in vitro assay for HDAC activity indicated that GhHDA 5 primarily deacetylates H3K9 acetylation marks. Moreover, the reduced expression of GhHDA 5 also suppresses fiber initiation and lint yield in the RNA interference (RNA i) lines. The 0 DPA ovules of GhHDA 5 RNA i lines also showed alterations in reactive oxygen species homeostasis and elevated autophagic cell death in the developing fibers. The differentially expressed genes (DEG s) identified through RNA ‐seq of RNA i line (DEP 12) and their pathway analysis showed that GhHDA 5 modulates expression of many stress and development‐related genes involved in fiber development. The reduced expression of GhHDA 5 in the RNA i lines also resulted in H3K9 hyper‐acetylation on the promoter region of few DEG s assessed by chromatin immunoprecipitation assay. The positively co‐expressed genes with GhHDA 5 showed cumulative higher expression during fiber initiation, and gene ontology annotation suggests their involvement in fiber development. Furthermore, the predicted protein interaction network in the positively co‐expressed genes indicates HDA 5 modulates fiber initiation‐specific gene expression through a complex involving reported repressors.     

Restoration of a Danube floodplain forest: what happens to species richness of terrestrial beetles?

Along the upper Danube, between river kilometer 2,472 and 2,464 (Bavaria, Germany), a managed hardwood forest was reconnected to the river via a newly carved floodplain channel. We report the stepwise alteration of the diversity of terrestrial beetles for six successive years from 2007 to 2012. In a 2‐year preliminary period (2007–2008), we recorded the baseline stage before the technical measures were implemented (2009–2010) and the onset of restoration occurred (2011–2012) with a continuous water flow in the new channel and seven flooding events. Each sample plot was equipped with a pitfall trap, an emergence photo‐eclector, an arboreal photo‐eclector, and a flight interception trap in breast height and in the canopy, respectively. The beetle communities act as an indicator to detect possible disturbance events when a riparian hardwood forest is stepwise transformed to become a new floodplain ecosystem. Within the 6‐year study period, we trapped 62,107 individual beetles, representing 85 families, 544 genera, and 1,191 species. Compared to the baseline stage, the abundance and the number of species decreased, including rare and red list species. On functional level, the species decline was particularly pronounced for zoophagous and mycetophagous species. Finally, we suppose that the 2‐year period since the launch of the new channel is too short for the establishment of a beetle community adjusted to the terrestrial part of the developing new floodplain forest.     

基于mRNA 5'端TIR区二级结构优化提高重组sTNFαRI在大肠杆菌中的表达水平

目的:通过优化PET11b-s TNFαRI 5'mRNA翻译起始区(TIR)二级结构从而提高可溶性肿瘤坏死因子I型受体(sTNFαRI)在大肠杆菌[E.coli BL21(DE3)]中的表达水平。方法:通过对PET11b-s TNFαRI mRNA 5'端TIR区二级结构的自由能及核苷酸位置熵分析,设计相应的引物对mRNA 5'翻译起始区(TIR)相应密码子进行突变,从而使核糖体结合位点(RBS)及起始密码子(AUG)暴露于发夹结构之外,此外将p ET11b核糖体结合位点由GAAGGAGA突变为GAAGAA,以利于翻译复合体的组装以及翻译起始。通过基因克隆的方法将5'端TIR区优化后的序列与s TNFαRI序列一起克隆到p ET11b载体中,并转化大肠杆菌BL21(DE3),阳性转化子经IPTG诱导表达,SDS-PAGE和Western blot检测。结果:通过对PET11b-s TNFαRI 5'TIR mRNA二级结构优化,经SDS-PAGE和Western blot分析表明重组s TNFαRI的表达水平较优化前提高50%~60%。结论:通过对重组载体翻译起始区(TIR)mRNA序列的二级结构优化可以有效提高目的蛋白的表达水平,对进一步工业化生产具有重要的应用价值。