Structural and functional characterization of the bacterial Cap3 enzyme in deconjugation and regulation of the cyclic dinucleotide transferase CD-NTase.

The Cyclic GMP-AMP synthase (cGAS) and cGAS/DncV-like nucleotidyltransferase (CD-NTase) enzymes belong to the key components of the innate immune sensor system that generates cyclic dinucleotide molecules in response to danger signals. Recently, it was discovered that CD-NTase in bacteria can undergo conjugation to protein substrates via an E1/E2 enzyme-mediated process, resembling ubiquitin modification system. Subsequently, these CD-NTase conjugated molecules will be hydrolyzed by the Cap3 enzyme in the same gene cluster. However, the experimental structure of bacterial CD-NTase recognized by Cap3 is unknown. Here, we first determined the crystal structure of the Cap3 enzyme in complex with the C-terminal tail of CD-NTase. Our structural and enzymatic analysis revealed that the C-terminal tail of CD-NTase is both necessary and sufficient for the Cap3-mediated hydrolysis of CD-NTase from its substrates. Interestingly, we further observed that after the hydrolysis reaction, the terminal glycine residue of the CD-NTase C-terminal tail was sequentially removed by Cap3, indicating that Cap3 might play a role in quenching the CD-NTase conjugation reaction. Our work provides experimental evidence elucidating the interaction between Cap3 and CD-NTase, and suggests a potential role for Cap3 in the bacterial Cyclic-oligonucleotide-based anti-phage signaling system (CBASS).

Active binding sites for ofloxacin resulted from adsorptive fractionation of humic acid on kaolinite.

The adsorptive fractionation of humic acid (HA) at the interface between minerals and water can significantly affect the fate of pollutants in water-soil environment. However, the adsorptive fractionation behavior of HA on kaolinite and its effect on the migration of fluoroquinolones (FQs) have not been fully understood. In this study, fluorescence and infrared spectroscopy, combined with two-dimensional correlation analyses, were used to explore the adsorptive fractionation of humic acid (HA) and its effects on ofloxacin adsorption on kaolinite. The results indicated that humic-like, rather than reduced quinone-like and tyrosine-like, was the main adsorptive fractionation component and preferentially bound to the Al-O sites of kaolinite. The adsorption mechanisms of humic-like and tyrosine-like mainly include hydrogen bonds between acidic functional groups and the Si-O or Al-O groups of kaolinite, n-π electron donor-acceptor interaction and electrostatic attraction. At pH 7.0, with addition of 4.0 and 16.0 mg C/L HA in solution, the adsorptive fractionation of HA on kaolinite led to increases in ofloxacin (in zwitterionic form) adsorption capacity by 1.46 and 3.35 mg/g, respectively. The interactions between ofloxacin and the humic-like were mainly hydrogen bonds and electrostatic attraction. Therefore, the influence of adsorptive fractionation of dissolved organic matter on minerals should be considered in estimating FQs environmental behaviors.

Differential compartmentalization of memory B cells versus plasma cells in salmonid fish.

The disposition of teleost memory and plasma cells (PCs) has essentially been un-explored. As the organization of the teleost immune system differs significantly from that of mammals (i.e. no bone marrow or lymph nodes, hematopoietic anterior kidney), this disposition could be essential in understanding how comparable functions are achieved. To address this question, the primary and secondary antibody-secreting cell, B memory cell, and antibody responses to T-independent and T-dependent antigens were analyzed in trout. Although the TI and TD antibody responses did not differ substantively from one another, the secondary responses to both were significantly prolonged compared with the primary responses. Logarithmic increases in titer and affinity were achieved for both antigens during the primary, with only modest increases during the secondary response. Antibody-secreting cells, both PCs and plasmablasts, quantitatively paralleled antibody production, with antibody-secreting cells skewing to the hematopoietic anterior kidney for both antigens. However, the enhanced antigen-inducible response of immune fish (indicative of the memory pool) skewed to the peripheral blood and spleen. This pattern of memory versus PC disposition parallels that observed in mammals even though the organization of the respective immune systems differs considerably.

The teleost humoral immune response.

Over the past 10 years our knowledge of cellular and molecular dynamics of teleost humoral immunity has increased enormously to now include: the existence of multiple isotypes, affinity-driven modulation of antibody structure and function, the unique trafficking patterns of each stage of B cell differentiation (including the plasma blast, short-lived and long-lived plasma cell, and the memory cell). Unfortunately the work which has generated the bulk of this information has generally employed defined antigens rather than vaccines. Thus, the focus of this review is to relate these aspects of immunity that are requisite for a mechanistic understanding of the generation of prophylactic immunity to the necessary analysis of responses to vaccines and vaccine candidates.

Alpha-2-Heremans-Schmid-glycoprotein (AHSG) a potential biomarker associated with prognosis of chromophobe renal cell carcinoma: The PROPOLIS study.

Background and Aims: Chromophobe renal cell carcinoma (chRCC) is the third common pathological subtype in renal cancers. However, the underlying mechanisms of specific genetic characteristics of chRCC are currently unclear. In this study, protein expression profiles, gene ontology (GO), and survival plots were provided by integrated bioinformatics analysis to investigate key genes associated with the mechanism of tumorigenesis and prognosis of chRCC. Methods: The chRCC data set of gene expression profiles and clinical data were obtained from the gdc-client (https://portal.gdc.cancer.gov) deposited on The Cancer Genome Atlas (TCGA) data portal. Differentially expressed genes (DEGs) in chRCC, compared with normal samples, were analyzed by R packages "DESeq2," "edgeR," and "limma." Heat maps, volcano plots, and principal component analysis (PCA) were performed for integrated analyses. GUniGO, mutant analysis, and survival plots were performed by R packages. A protein-protein interaction (PPI) network was generated and analyzed by R packages, online String software, and Cytoscape software. Survival analysis and gene expressing comparison in tumor and normal samples were used to detect the core genes of chRCC. Furthermore, the top interacting proteins were reanalyzed. Results: A total of 306 upregulated genes and 678 downregulated genes were identified by a Venn diagram. Ten hub genes were extracted from PPI network. Furthermore, Alpha-2-Heremans-Schmid-glycoprotein (AHSG), one of 10 hub genes, was found to be associated with chRCC, and had a big difference in expression between survival and dead events. AHSG could predict potential prognostic and may be a diagnostic biomarker in chRCC. Conclusion: This study illustrated that AHSG may be a potential therapeutic target and prognostic genetic marker for chRCC.

White spot syndrome virus VP37 interacts with VP28 and VP26.

White spot syndrome virus (WSSV) is one of the most virulent pathogens affecting penaeid shrimp, causing high mortality in infected populations. Interactions between virus structural proteins are likely to be important for virus assembly. Many steps of the WSSV assembly and maturation pathway remain unclear. In the present study, the interaction between VP37 and envelope or nucleocapsid proteins was characterized. VP37 was expressed in Escherichia coli and confirmed by Western blotting. Pure WSSV virions were subjected to Triton X-100 treatment to separate the envelope and nucleocapsid fractions. Overlay assays showed that VP37 interacted with VP28 and VP26. The interaction of VP37 with VP28 and VP26 was confirmed further by His pull-down and matrix-assisted laser desorption ionization (MALDI) mass spectrographic assays. The binding assay of VP37 with VP28 by ELISA confirmed that the 2 proteins had direct interaction in vivo. This discovery will help elucidate the molecular mechanisms of virion morphogenesis.