Identification of a virus trans-acting regulatory element on the latent DNA replication of Kaposi's sarcoma-associated herpesvirus

Latency-associated nuclear antigen 1 (LANA1) of Kaposi's sarcoma-associated herpesvirus (KSHV) plays a pivotal role in the maintenance of the virus genome in latently infected cells. LANA1 links virus genomes to host chromosomes via a C-terminal DNA-binding domain which interacts with the sequences located in terminal repeats (TRs) of the virus genome and via an N-terminal chromosome-binding sequence which associates with the host chromosomes, respectively. Recent data suggest that LANA1 also actively participates in the replication of KSHV TR-containing plasmid in the transient DNA replication assay. In this report, it was found that C33A and COS-1, but not NIH/3T3, cell lines are permissive for the transient replication of KSHV TR-containing plasmid. Using several LANA1-deletion mutants, the minimum domain of LANA1 required for replication activity was also determined. In addition, the N terminus of LANA1 inhibited the transient replication systems of KSHV and Epstein-Barr virus (EBV) in transiently transfected 293 and 293T cells, but the C terminus of LANA1 specifically inhibited the transient replication system of KSHV in other cell lines. Consistent with previous reports, these data further emphasize the functional importance of the N terminus of LANA1 on replication from the KSHV latent origin of DNA replication.close141

Inhibition of nuclear factor kappa B activity by viral interferon regulatory factor 3 of Kaposi's sarcoma-associated herpesvirus

Nuclear factor-kappaB (NF-kappaB) is a transcription factor that plays an important role in the immune system and cell death. Many viral proteins modulate NF-kappaB to escape host immune surveillance, promote cell survival, and enhance viral replication. In the present study, we show that NF-kappaB activity is downmodulated by viral interferon regulatory factor 3 (vIRF3), which is encoded by Kaposi's sarcoma-associated herpesvirus open-reading frame K10.5. vIRF3 repressed NF-kappaB-dependent transcription in a dose-dependent manner and inhibited the activation of NF-kappaB induced by tumor necrosis factor (TNF)-alpha. In vivo studies showed vIRF3 inhibited IkappaB kinase beta (IKKbeta) activity, but not IKKalpha activity, resulting in reduced IkappaB phosphorylation. Immunofluorescence assays showed that vIRF3 interfered with nuclear translocation of NF-kappaB. In addition, consistent with the inhibition of NF-kappaB activity, vIRF3 sensitized cells to TNF-alpha-induced apoptosis. While vIRF3 interacts with IKKbeta in vitro and in 293T cells, we were unable to demonstrate vIRF3-IKKbeta interaction in BCBL-1 cells. Our results indicate that vIRF3 can regulate the host immune system and apoptosis via inhibition of NF-kappaB activity.close383

Sumoylation of the Novel Protein hRIPβ Is Involved in Replication Protein A Deposition in PML Nuclear Bodies

Replication protein A (RPA) is a single-stranded-DNA-binding protein composed of three subunits with molecular masses of 70, 32, and 14 kDa. The protein is involved in multiple processes of eukaryotic DNA metabolism, including DNA replication, repair, and recombination. In Xenopus, Xenopus RPA-interacting protein α has been identified as a carrier molecule of RPA into the nucleus. In this study, human RPA-interacting protein α (hRIPα) and five novel splice isoforms (named hRIPα, hRIPβ, hRIPγ, hRIPδ1, hRIPδ2, and hRIPδ3 according to the lengths of their encoding peptides) were cloned. Among hRIP isoforms, hRIPα and hRIPβ were found to be the major splice isoforms and to show different subcellular localizations. While hRIPα localized to the cytoplasm, hRIPβ was found in the PML nuclear body. Modification of hRIPβ by sumoylation was found to be required for localization to the PML nuclear body. The results of the present work demonstrate that hRIPβ transports RPA into the PML nuclear body and releases RPA upon UV irradiation. hRIPβ thus plays an important role in RPA deposition in PML nuclear bodies and thereby supplements RPA for DNA metabolism

Lysobacter humi sp. nov., isolated from soil

Lee, Dongwook, Jang, Jun Hyeong, Cha, Seho, Seo, Taegun (2017): Lysobacter humi sp. nov., isolated from soil. International Journal of Systematic and Evolutionary Microbiology 67 (4): 951-955, DOI: 10.1099/ijsem.0.001722, URL: http://dx.doi.org/10.1099/ijsem.0.00172

Characteristics and Biological Activity of Exopolysaccharide Produced by <i>Lysobacter</i> sp. MMG2 Isolated from the Roots of <i>Tagetes patula</i>

In the present study, exopolysaccharide (EPS) produced by Lysobacter sp. MMG2 (lyEPS) was characterized and purified. The lyEPS-producing strain Lysobacter sp. MMG2 was isolated from the roots of Tagetes patula. When lyEPS was produced in tryptic soy broth with 1% glucose and the lyophilized powder was measured, the yield was found to be 0.67 g/L. The molecular weight (Mw) of lyEPS was 1.01 × 105 Da. Its monosaccharide composition includes 84.24% mannose, 9.73% glucose, 2.55% galactose, 2.77% arabinose, 0.32% xylose, and 0.03% rhamnose. Scanning electron microscopy (SEM) revealed that lyEPS has various round and rough surfaces. Fourier-transform infrared (FTIR) analysis identified its carbohydrate polymer functional groups. Moreover, thermogravimetric analysis of lyEPS revealed two events of mass loss: the first was water loss, which resulted in 3.97% mass loss and the second event occurred at approximately 212 °C. lyEPS could inhibit biofilm-producing pathogenic bacteria without any antimicrobial activity. Furthermore, lyEPS at a concentration of 4 mg/mL could exhibit potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity (89.25%). These results indicate that lyEPS could be a promising candidate for industrial development if its biological activity is further explored

An Isolated Arthrobacter sp. Enhances Rice (Oryza sativa L.) Plant Growth

Rice is a symbol of life and a representation of prosperity in South Korea. However, studies on the diversity of the bacterial communities in the rhizosphere of rice plants are limited. In this study, four bundles of root samples were collected from the same rice field located in Goyang, South Korea. These were systematically analyzed to discover the diversity of culturable bacterial communities through culture-dependent methods. A total of 504 culturable bacteria were isolated and evaluated for their plant growth-promoting abilities in vitro. Among them, Arthrobacter sp. GN70 was selected for inoculation into the rice plants under laboratory and greenhouse conditions. The results showed a significantly positive effect on shoot length, root length, fresh plant weight, and dry plant weight. Moreover, scanning electron microscopic (SEM) images demonstrated the accumulation of bacterial biofilm networks at the junction of the primary roots, confirming the root-colonizing ability of the bacterium. The strain also exhibited a broad spectrum of in vitro antimicrobial activities against bacteria and fungi. Here, we first report the rice plant growth-promoting ability of the Arthrobacter species with the biofilm-producing and antimicrobial activities against plant and human pathogens. Genome analyses revealed features attributable to enhance rice plant growth, including the genes involved in the synthesis of plant hormones, biofilm production, and secondary metabolites. This study revealed that the rhizobacteria isolated from the roots of rice plants have dual potential to be utilized as a plant growth promoter and antimicrobial agent

DNA-PK/Ku complex binds to latency-associated nuclear antigen and negatively regulates Kaposi&apos;s sarcoma-associated herpesvirus latent replication

During latent infection, latency-associated nuclear antigen (LANA) of Kaposi&apos;s sarcoma-associated herpesvirus (KSHV) plays important roles in episomal persistence and replication. Several host factors are associated with KSHV latent replication. Here, we show that the catalytic subunit of DNA protein kinase (DNA-PKcs), Ku70, and Ku86 bind the N-terminal region of LANA. LANA was phosphorylated by DNA-PK and overexpression of Ku70, but not Ku86, impaired transient replication. The efficiency of transient replication was significantly increased in the HCT116 (Ku86 +/-) cell line, compared to the HCT116 (Ku86 +/+) cell line, suggesting that the DNA-PK/Ku complex negatively regulates KSHV latent replication.close7

Viral Interferon Regulatory Factor 1 of Kaposi's Sarcoma-Associated Herpesvirus Interacts with a Cell Death Regulator, GRIM19, and Inhibits Interferon/Retinoic Acid-Induced Cell Death

Kaposi's sarcoma-associated herpesvirus (KSHV) plays a significant role in the development of Kaposi's sarcoma, primary effusion lymphoma, and some forms of multicentric Castleman's disease. The KSHV open reading frame K9 encodes the viral interferon (IFN) factor 1 (vIRF1), which downregulates IFN- and IRF-mediated transcriptional activation, and leads to cellular transformation in rodent fibroblasts and induction of tumors in nude mice. Using the yeast two-hybrid assay, we identified genes associated with retinoid-IFN-induced mortality-19 (GRIM19), which interacts directly with vIRF1, both in vivo and in vitro. The N-terminal region of vIRF1 is required for binding GRIM19. Colocalization of vIRF1 and GRIM19 was observed in 293T cells. The vIRF1 protein deregulates GRIM19-induced apoptosis in the presence of IFN/all-trans-retinoic acid (RA) and inhibits IFN/RA-induced cell death. Another DNA tumor viral protein, human papillomavirus type 16 E6, also binds GRIM19, suggesting that this is a general target of viral proteins. Our results collectively indicate that vIRF1 modulates IFN/RA-cell death signals via interactions with GRIM19