Novel variant of p230 trans-Golgi network protein identified by serum from Sjögren's syndrome patient

Trans-Golgi network (TGN) protein p230 is a peripheral membrane protein associated with the cytoplasmic face of the TGN. TGNp230 is an extensively coiled-coil protein with flexible amino- and carboxyl-terminal ends, associates with non-clathrin-coated vesicles arising from the TGN, and is implicated in vesicle biogenesis. Here we used an autoimmune serum from a patient with S ogren's syndrome to clone partial cDNAs from a human hepatoma HepG2 expression library. The partial cDNAs encoded a novel amino-terminal splice variant of TGNp230. Specific reactivity of the autoimmune serum for p230 is supported by immunofluorescene staining of the Golgi apparatus, immunoblotting of a > 200-kDa HeLa cell protein, and reactivity with a bacterially expressed GST-p230 fusion protein. The alternative splicing occurs within the first proline-rich domain of p230. It comprises a deletion of 30 bp followed immediately by an additional 66 bp absent in the published sequence. RT-PCR analysis indicated that the splicing occurs independently of previously reported carboxyl-terminal splicing, and that this novel splice variant is more frequent than the previously reported p230. The novel splice variant of p230 is also located at the TGN. We propose that p230 splice variants may be implicated in selection of cargo molecules for vesicles arising from the TGN.     

ATL3 Is a Tubular ER-Phagy Receptor for GABARAP-Mediated Selective Autophagy

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枯草芽孢杆菌中一种新型双向启动子的功能鉴定

本研究旨在通过转录组分析预测的方法,由地衣芽孢杆菌中筛选获得一种新型双向启动子,鉴定其启动强度。以已知强组成型启动子pShuttle-09为对照,检测其对克劳氏芽孢杆菌碱性蛋白酶基因的表达活性。成功构建了3种重组碱性蛋白酶表达载体及对应的工程菌株。在新型启动子pLA和其反向启动子pLB调控转录下,克劳氏芽孢杆菌碱性蛋白酶表达活性达到164 U/mL和111 U/mL。结果表明,pLA的启动强度明显高于pShuttle-09和pLB,pLA启动子与pLB启动子均可表达碱性蛋白酶。从而为枯草芽孢杆菌表达系统中异源基因的表达提供一个新的方向,也为原核生物中共同表达两种基因提供了新的思路。     

Identification and characterization of a novel cold-tolerant extracellular protease from Planococcus sp. CGMCC 8088

Extremophiles - A psychrophilic extracellular protease was isolated from the marine bacterium Planococcus sp. M7 found in the deep-sea mud of the Southern Indian Ocean. The mature protease is about...     

Structural Variation and Uniformity among Tetraloop-Receptor Interactions and Other Loop-Helix Interactions in RNA Crystal Structures

Tetraloop-receptor interactions are prevalent structural units in RNAs, and include the GAAA/11-nt and GNRA-minor groove interactions. In this study, we have compiled a set of 78 nonredundant loop-helix interactions from X-ray crystal structures, and examined them for the extent of their sequence and structural variation. Of the 78 interactions in the set, only four were classical GAAA/11-nt motifs, while over half (48) were GNRA-minor groove interactions. The GNRA-minor groove interactions were not a homogeneous set, but were divided into five subclasses. The most predominant subclass is characterized by two triple base pair interactions in the minor groove, flanked by two ribose zipper contacts. This geometry may be considered the “standard” GNRA-minor groove interaction, while the other four subclasses are alternative ways to form interfaces between a minor groove and tetraloop. The remaining 26 structures in the set of 78 have loops interacting with mostly idiosyncratic receptors. Among the entire set, a number of sequence-structure correlations can be identified, which may be used as initial hypotheses in predicting three-dimensional structures from primary sequences. Conversely, other sequence patterns are not predictive; for example, GAAA loop sequences and GG/CC receptors bind to each other with three distinct geometries. Finally, we observe an example of structural evolution in group II introns, in which loop-receptor motifs are substituted for each other while maintaining the larger three-dimensional geometry. Overall, the study gives a more complete view of RNA loop-helix interactions that exist in nature.     

Chemical Constituents from a Mangrove-Derived Actinobacteria Isoptericola chiayiensis BCRC 16888 and Evaluation of Their Anti-NO Activity

Cultivation of the actinobacteria strain Isoptericola chiayiensis, a mangrove - derived actinobacteria that was isolated from a mangrove soil collected in Chiayi County, resulted in the isolation of one new 2-furanone derivative, isopterfuranone ( 1 ), one new sesquiterpenoid, isopterchiayione ( 2 ), one new benzenoid derivative, isopterinoid ( 3 ), five new flavonoids, chiayiflavans A−E ( 4 – 8 ), and 4 metabolites isolated for the first time from nature source, methyl 3-(4-methyl-2,5-dioxopyrrolidin-3-yl)propanoate ( 9 ), 3-ethyl-4-methylpyrrolidine-2,5-dione ( 10 ), chiayiensol ( 11 ) and chiayiensic acid ( 12 ). Their structures were determined through in-depth spectroscopic and mass-spectrometric analyses. Most of the isolates showed potent inhibitory effects on NO production in LPS-stimulated RAW 264.7 murine macrophages cells with IC₅₀ values ranging from 9.36 to 40.02 μM. Of these isolates, 4 and 5 showed NO inhibitory activity with IC₅₀ values of 17.14 and 9.36 μM, stronger than the positive control quercetin (IC₅₀=36.95 μM). This is the first report on flavan metabolites from the genus Isoptericola.     

Segregation of Deoxyribonucleic Acid in Bacteria: Association of the Segregating Unit with the Cell Envelope

Cells of the gram-positive organism Lactobacillus acidophilus R-26 were labeled with ³H-thymine to measure the segregation of radioactive deoxyribonucleic acid (DNA) into daugher cells. Such cells were found to contain 8 conserved units of DNA which would correspond to two replicating chromosomes per cell. Fluorescent antibody (FA) against this organism was used to demonstrate that portions of the cell surface (2 to 4 units per cell) were conserved during growth and division. The permanent association of DNA with these conserved cell surface units was measured by combining autoradiography with FA techniques. DNA synthesized immediately before FA labeling was not associated with the fluorescent cell surface, whereas DNA synthesized a generation previously was. The results are consistent with a model in which DNA becomes permanently fixed to the cell surface when it is first used as a template.     

A Bacillus subtilis sensor kinase involved in triggering biofilm formation on the roots of tomato plants

The soil bacterium Bacillus subtilis is widely used in agriculture as a biocontrol agent able to protect plants from a variety of pathogens. Protection is thought to involve the formation of bacterial communities - biofilms - on the roots of the plants. Here we used confocal microscopy to visualize biofilms on the surface of the roots of tomato seedlings and demonstrated that biofilm formation requires genes governing the production of the extracellular matrix that holds cells together. We further show that biofilm formation was dependent on the sensor histidine kinase KinD and in particular on an extracellular CACHE domain implicated in small molecule sensing. Finally, we report that exudates of tomato roots strongly stimulated biofilm formation ex planta and that an abundant small molecule in the exudates, (L) -malic acid, was able to stimulate biofilm formation at high concentrations in a manner that depended on the KinD CACHE domain. We propose that small signalling molecules released by the roots of tomato plants are directly or indirectly recognized by KinD, triggering biofilm formation.     

Green synthesis of silver nanoparticles using Rhodobacter Sphaeroides

The use of microorganisms in the synthesis of nanoparticles emerges as an eco-friendly and exciting approach. In this study, silver nanoparticles were successfully synthesized from AgNO₃ by reduction of aqueous Ag⁺ ions with the cell filtrate of Rhodobacter sphaeroides. Nanoparticles were characterized by means of UV–vis absorption spectroscopy, X-Ray Diffraction (XRD), transmission electron microscopy (TEM) and high-resolution transmission electron microscopy (HRTEM). Crystalline nature of the nanoparticles in the fcc structure are confirmed by the peaks in the XRD pattern corresponding to (111), (200), (220) and (311) planes, bright circular spots in the selected are a electron diffraction (SAED) and clear lattice fringes in the high-resolution TEM image. Also, the size of silver nanoparticles was controlled by the specific activity of nitrate reductase in the cell filtrate.