Mitochondrial gene rearrangement and phylogenetic relationships in the Amphilepidida and Ophiacanthida (Echinodermata, Ophiuroidea)

Mitochondrial genomes provide an additional data source for nuclear phylogenomics. Recently, the taxonomic classification of Ophiuroidea was changed dramatically based on a molecular phylogenetic analysis that utilized a huge nuclear dataset of transcriptome and target-capture approaches. However, the mitochondrial genome analysis of Ophiuroidea was not conducted sufficiently in depth to allow comparison with current phylogenetic relationships, especially for the Ophintegrida. In this study, eight mitogenomes were newly reported in Amphilepidida and Ophiacanthida. Phylogenetic analyses were undertaken based on the nucleotide sequences of 13 protein coding genes (PCGs), 22 tRNA, and two rRNA, and the amino acid translated sequences of 13 PCGs. The results of the phylogenetic analysis suggested that the amino acid translated sequences were more useful than the nucleotide sequences for the analysis of higher phylogenetic relationships. The mitochondrial gene order of 13 PCGs and two rRNA in Amphilepidida and Ophiacanthida was relatively conserved. However, more complex gene orders occurred in the phylogeny of Amphilepidida and Ophiacanthida, including 22 tRNA. The results of our study suggest that Ophiuroidea has undergone more complex gene rearrangements than other classes of echinoderms.</p

Activation of the p21 promoter by JMJD2D in HEK293T cells.

<p>(<b>A</b>) Activity of a p21 luciferase reporter construct upon cotransfection of control vector pEV3S or wild-type or H192A JMJD2D is depicted. As indicated, empty vector pcDNA3 or pcDNA3-p53 was also transfected. (<b>B</b>) Analogous, response of a CMV or MMP-1 luciferase reporter construct to cotransfection of p53 and/or JMJD2D. (<b>C</b>) Chromatin immunoprecipitation assay in HCT116 cells treated without and with adriamycin.</p

Mapping of interaction domains.

<p>(<b>A</b>) Indicated Flag-tagged amino acids of JMJD2D were coexpressed with p53 in HEK293T cells and complex formation assessed in coimmunoprecipitation assays as in Fig. 1B. A sketch of human JMJD2D highlighting its JmjC domain is presented at the bottom. (<b>B</b>) Comparable amounts of GST or indicated GST-p53 fusion proteins were bound to glutathione agarose. After incubation with Flag-tagged JMJD2D, bound JMJD2D was revealed by anti-Flag Western blotting. The location of the DNA binding domain within p53 is indicated in the sketch at the bottom.</p

JMJD2D protects from apoptosis.

<p>(<b>A</b>) The level of sub-G1 HCT116 cells was determined in cells that expressed control shRNA or JMJD2D shRNA#3. Cells were treated for 72 h with 1 µM adriamycin or DMSO as indicated. (<b>B</b>) The same in case of p53^−/− HCT116 cells. (<b>C</b>) Model of JMJD2D action.</p

Binding of JMJD2D to p53.

<p>(<b>A</b>) Flag-tagged JMJD2D or HSPBAP1 were coexpressed with p53 in HEK293T cells. After anti-p53 immunoprecipitation (IP), coprecipitated proteins were revealed by anti-Flag immunoblotting (top panel). The bottom two panels show input levels of Flag-tagged proteins or p53. IgH, immunoglobulin heavy chain. (<b>B</b>) Indicated Flag-tagged JMJD proteins were coexpressed with p53 in HEK293T cells. After anti-Flag immunoprecipitation, coprecipitated p53 was detected by anti-p53 Western blotting (top panel). The middle and bottom panels show p53 and Flag-JMJD protein input levels, respectively. (<b>C</b>) HCT116 cell extracts were challenged with no, control or anti-JMJD2D antibodies and coprecipitated p53 detected by immunoblotting (top panel). The bottom panel shows that respective JMJD2D input levels were equal.</p

Stimulation of p21 expression by JMJD2D in U2OS cells.

<p>(<b>A</b>) Wild-type or H192A JMJD2D or empty vector pEV3S were stably transfected into U2OS cells. Expression of indicated proteins was assessed by Western blotting in cells treated for 24 h with 1 µM adriamycin or DMSO as a control. (<b>B</b>) RNA was isolated from stably transfected U2OS cells and RT-PCR analyses were performed. Shown is the amplification of p21 cDNA and, as a control, GAPDH cDNA.</p

JMJD2D depletion results in reduced cell proliferation.

<p>(<b>A</b>) HCT116 cells expressing control or JMJD2D shRNA were treated with 1 µM adriamycin or with DMSO as a control for 24 h. Downregulation of JMJD2D was assessed by Western blotting; immunoblotting for actin served as a loading control. (<b>B</b>) Wild-type or p53^−/− HCT116 cells were challenged with control or JMJD2D shRNA (#1 or #3) and then treated without or with 1 µM adriamycin for 72 h. The number of cells were counted and presented as percent of the control shRNA for each wild-type and p53^−/− HCT116 cells. Statistical significance of differences between various experimental conditions is indicated in the graph.</p

DataSheet_1_Methylation of the epigenetic JMJD2D protein by SET7/9 promotes prostate tumorigenesis.pdf

How the function of the JMJD2D epigenetic regulator is regulated or whether it plays a role in prostate cancer has remained elusive. We found that JMJD2D was overexpressed in prostate tumors, stimulated prostate cancer cell growth and became methylated by SET7/9 on K427. Mutation of this lysine residue in JMJD2D reduced the ability of DU145 prostate cancer cells to grow, invade and form tumors and elicited extensive transcriptomic changes. This included downregulation of CBLC, a ubiquitin ligase gene with hitherto unknown functions in prostate cancer, and upregulation of PLAGL1, a transcription factor with reported tumor suppressive characteristics in the prostate. Bioinformatic analyses indicated that CBLC expression was elevated in prostate tumors. Further, downregulation of CBLC largely phenocopied the effects of the K427 mutation on DU145 cells. In sum, these data have unveiled a novel mode of regulation of JMJD2D through lysine methylation, illustrated how this can affect oncogenic properties by influencing expression of the CBLC gene, and established a pro-tumorigenic role for CBLC in the prostate. A corollary is that JMJD2D and CBLC inhibitors could have therapeutic benefits in the treatment of prostate and possibly other cancers.</p

Anti-inflammatory Amino Acid Derivatives from the Ascidian <i>Herdmania momus</i>

Four new amino acid derivatives, herdmanines A–D (1–4), were isolated from the marine ascidian Herdmania momus. Herdmanines A–C contain the unusual d-form of arginine. Compounds 3 and 4 had a moderate suppressive effect on the production of NO, with IC50 values of 96 and 9 μM, respectively. These compounds were found to inhibit the mRNA expression of iNOS. The inhibitory activities on the production and mRNA expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 were evaluated

Additional file 1 of Distribution and antimicrobial resistance profiles of bacterial species in stray cats, hospital-admitted cats, and veterinary staff in South Korea

Additional file 1 Figure S1. Percentage resistance for antimicrobials against Coagulase-positive Staphylococci (A), Coagulase-negative Staphylococci (B), Enterobacteriaceae (C), and Enterococcus spp. (D) isolates from stray cats, hospital-admitted cats, and veterinary staff. The data sets labeled with different superscript letters (a, b, and c) are statistically different from each other (P < 0.05)