RESUMO
Purified translesion synthesis (TLS) DNA polymerases (Pols) replicate through DNA lesions with a low fidelity; however, TLS operates in a predominantly error-free manner in normal human cells. To explain this incongruity, here we determine whether Y family Pols, which play an eminent role in replication through a diversity of DNA lesions, are incorporated into a multiprotein ensemble and whether the intrinsically high error rate of the TLS Pol is ameliorated by the components in the ensemble. To this end, we provide evidence for an indispensable role of Werner syndrome protein (WRN) and WRN-interacting protein 1 (WRNIP1) in Rev1-dependent TLS by Y family Polη, Polι, or Polκ and show that WRN, WRNIP1, and Rev1 assemble together with Y family Pols in response to DNA damage. Importantly, we identify a crucial role of WRN's 3' â 5' exonuclease activity in imparting high fidelity on TLS by Y family Pols in human cells, as the Y family Pols that accomplish TLS in an error-free manner manifest high mutagenicity in the absence of WRN's exonuclease function. Thus, by enforcing high fidelity on TLS Pols, TLS mechanisms have been adapted to safeguard against genome instability and tumorigenesis.
Assuntos
DNA Polimerase Dirigida por DNA , Síntese de DNA Translesão , Helicase da Síndrome de Werner , Humanos , Dano ao DNA , Reparo do DNA , Replicação do DNA , DNA Polimerase Dirigida por DNA/metabolismo , Exonucleases/metabolismo , Síntese de DNA Translesão/genética , Helicase da Síndrome de Werner/genética , Helicase da Síndrome de Werner/metabolismoRESUMO
A Gram-stain-negative, aerobic, rod-shaped, motile, flagellated bacterial strain, designated as CAU 1639T, was isolated from the tidal flat sediment on the Yellow Sea in the Republic of Korea. Growth of the isolate was observed at 20-37 °C, at pH 5.0-10.5 and with 0-7% (w/v) NaCl. The genomic DNA G + C content was 60.8%. Phylogenetic analysis, grounded on 16S rRNA gene sequencing, revealed that strain CAU 1639T was closely related to species within the genus Roseibium. It shared the highest similarity with Roseibium album CECT 5095T, followed by Roseibium aggregatum IAM 12614T and Roseibium salinum Cs25T, with 16S rRNA gene sequence similarity ranging from 98.0-98.4%. It was observed that the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values ranged between 72.5-79.5 and 20.0-22.9%, respectively. The polyphasic taxonomic analysis reveals that strain CAU 1639T represents a novel species in the genus Roseibium with the proposed name Roseibium sediminicola sp. nov. The type strain is CAU 1639T (= KCTC 82430T = MCCC 1K06081T).
Assuntos
Composição de Bases , DNA Bacteriano , Sedimentos Geológicos , Filogenia , RNA Ribossômico 16S , Água do Mar , Sedimentos Geológicos/microbiologia , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , República da Coreia , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Rhodobacteraceae/classificação , Rhodobacteraceae/genética , Rhodobacteraceae/isolamento & purificação , Análise de Sequência de DNA , Hibridização de Ácido Nucleico , Ácidos Graxos/análise , Ácidos Graxos/química , DNA Ribossômico/genéticaRESUMO
A Gram-stain-negative, strictly aerobic, non-motile, rod-shaped, designated strain CAU 1642 T, was isolated from a Salicornia herbacea collected from a tidal flat in the Yellow Sea. Strain CAU 1642 T grew optimally at pH 8.0 and 30 °C. The highest 16S rRNA gene sequence similarity was 97.25%, with Pseudomarinomonas arenosa CAU 1598 T, and phylogenetic analysis indicated that strain CAU 1642 T belongs to the genus Pseudomarinomonas. The major cellular fatty acids were iso-C15:0, iso-C16:0, and summed feature 9 (iso-C17:1ω9c and/or 10-methyl C16:0). Ubiquinone-8 was the major respiratory quinone. The draft genome of strain CAU 1642 T was 4.5 Mb, with 68.7 mol% of G + C content. The phylogenetic, phenotypic, and chemotaxonomic analysis data reveal strain CAU 1642 T to be of a novel genus in the family Lysobacteraceae, with the proposed name Pseudomarinomonas salicorniae sp. nov. with type strain CAU 1642 T (= KCTC 92084 T = MCCC 1K07085T).
Assuntos
Composição de Bases , Chenopodiaceae , DNA Bacteriano , Ácidos Graxos , Filogenia , RNA Ribossômico 16S , Chenopodiaceae/microbiologia , RNA Ribossômico 16S/genética , Ácidos Graxos/análise , Ácidos Graxos/química , DNA Bacteriano/genética , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Análise de Sequência de DNA , Quinonas/análise , Ubiquinona/química , Ubiquinona/análogos & derivados , Genoma BacterianoRESUMO
A Gram-staining-positive, aerobic, non-spore-forming bacterium was isolated from coastal sand samples from Incheon in the Republic of Korea and designated as strain CAU 1645T. The optimum conditions for growth were observed at 30 °C in growth media containing 1% (w/v) NaCl at pH 9.0. The predominant respiratory quinone was MK-9 and the major fatty acids were C16:0, C17:1 w7c, and summed feature 7. Similarly, the 16S rRNA gene sequence exhibited the highest similarity with Mycolicibacterium bacteremicum DSM 45578T and Mycolicibacterium neoaurum JCM 6365T, both of which exhibited similarity rates of 97.2%. The genomic DNA G+C content was 68.2%. The whole genome of strain CAU 1645T was obtained and annotated with annotation using RAST server. The pan-genome analysis was determined using Prokka, Roary, and Phandango. In the pan-genome analysis, the strain CAU 1645T shared 40 core genes with closely related Mycolicibacterium species, including the AcpM gene, the meromycolate extension acyl carrier protein involved in forming impermeable cell walls in mycobacteria. Therefore, our findings demonstrated that the isolate represents a novel species of the genus Mycolicibacterium, for which we propose the name Mycolicibacterium arenosum sp. nov. The type strain is CAU 1645T (= KCTC 49724T = MCCC 1K07087T).
Assuntos
Proteína de Transporte de Acila , Areia , RNA Ribossômico 16S/genética , Parede Celular , Meios de CulturaRESUMO
A Gram-stain-negative, aerobic, non-flagellated and coccoid or ovoid bacterial strain, BSSL-BM11T, was isolated from sand of coastal dunes along the Yellow Sea of the Korean peninsula. Strain BSSL-BM11T grew optimally at 30â°C, at pH 7.0-8.0 and in the presence of 2.0-3.0â% (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences, the up-to-date bacterial core gene set and average amino acid identity (AAI) showed that strain BSSL-BM11T forms a cluster with the type strains of Tianweitania sediminis and Corticibacterium populi. Strain BSSL-BM11T showed 16S rRNA gene sequence similarities of 98.3 and 98.0â% to the type strains of T. sediminis and C. populi, respectively, and less than 96.4â% to the type strains of the other recognized species. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain BSSL-BM11T and the type strains of T. sediminis and C. populi were 77.0-84.8â% and 20.0-28.1â%, respectively. The 16S rRNA gene similarity, AAI, ANI and dDDH values between T. sediminis Z8T and C. populi KCTC 42249T were 98.0, 77.4, 76.7 and 20.1â%, respectively. The DNA G+C content of strain BSSL-BM11T from genomic sequence data was 61.3âmol%. Strain BSSL-BM11T contained Q-10 as the predominant ubiquinone and C18â:â1 ω7c, C16â:â0 and cyclo C19â:â0 ω8c as the major fatty acids. The major polar lipids of strain BSSL-BM11T were phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. Based on the polyphasic data, it is proposed that C, populi be reclassified as a member of the genus Tianweitania. Phenotypic and phylogenetic analyses revealed that strain BSSL-BM11T is separated from T. sediminis and C. populi. On the basis of the data presented here, strain BSSL-BM11T (=KACC 21634T=NBRC 114503T) is considered to represent a novel species of the genus Tianweitania, for which the name Tianweitania aestuarii sp. nov. is proposed.
Assuntos
Ácidos Graxos , Ubiquinona , Ácidos Graxos/química , Filogenia , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Análise de Sequência de DNA , Composição de Bases , Técnicas de Tipagem Bacteriana , Ubiquinona/química , Fosfolipídeos/químicaRESUMO
Atopic dermatitis (AD) is a chronic immune disease that requires long-term management owing to its relative ease of recurrence. However, steroid treatment is limited owing to the side effects. Therefore, research on therapeutics with proven safety is required. Here, we evaluated the anti-allergic activity of the probiotic strain Pediococcus pentosaceus KF159 (PPKF159) with an ex vivo mouse model sensitized with ovalbumin (OVA) and a mouse model of AD induced by house dust mites. Changes in pathological symptoms were confirmed based on the clinical status of the AD-induced lesion site and the levels of T helper type 2 (Th2)-derived cytokines and immunoglobulin E (IgE). In addition, cell-mediated responses and related mechanisms were elucidated using various kinds of primary cells including splenocytes, mesenteric lymph nodes, Peyer's patch, and bone marrow-derived dendritic cells (BMDCs) in vitro and ex vivo. Oral administration of PPKF159 alleviated AD-like clinical symptoms such as erythema, edema, hemorrhage, and increased tissue thickness, and suppressed the production of Th2-associated cytokines and serum IgE while increasing T helper type 1 (Th1)-mediated cytokine production. PPKF159 induced tolerogenic dendritic cells (tol-DCs) by increasing the expression of ICOS-L, PD-L1, and IDO which were closely related to Treg induction in PPKF159-treated BMDCs. In addition, BMDCs and naive T cells co-cultured in the presence of PPKF159 had elevated IL10 production and increased proportions of CD4+CD25+Foxp3+ Tregs compared to the absence of PPKF159. This study showed that PPKF159 relieved AD-like clinical symptoms, modulated the Th1/Th2 immune balance, and inhibited IgE production in a mouse AD model. PPKF159 induced the transformation of dendritic cells into tolerogenic versions. These induced tol-DCs directly enhanced the production of IL10 or improved the secretion of IL10 through the induction of CD4+CD25+Foxp3+ Treg cells, thereby improving AD. These results suggest that PPKF159 can be applied as a functional food material for the treatment and prevention of AD.