Expanding and understanding the genetic toolbox of the hyperthermophilic genus Sulfolobus

2009 ◽  
Vol 37 (1) ◽  
pp. 97-101 ◽  
Author(s):  
Michaela Wagner ◽  
Silvia Berkner ◽  
Malgorzata Ajon ◽  
Arnold J.M. Driessen ◽  
Georg Lipps ◽  
...  
Keyword(s):  
Deletion Mutant ◽  
Sulfolobus Solfataricus ◽  
Sulfolobus Acidocaldarius ◽  
Exogenous Dna ◽  
Targeted Deletion ◽  
Genetic Tools ◽  
Mutant Strains ◽  
Micro Organisms

Although Sulfolobus species are among the best studied archaeal micro-organisms, the development and availability of genetic tools has lagged behind. In the present paper, we discuss the latest progress in understanding recombination events of exogenous DNA into the chromosomes of Sulfolobus solfataricus and Sulfolobus acidocaldarius and their application in the construction of targeted-deletion mutant strains.

scholarly journals N-3-Hydroxy Dodecanoyl-DL-homoserine Lactone (OH-dDHL) Triggers Apoptosis of Bone Marrow-Derived Macrophages through the ER- and Mitochondria-Mediated Pathways

2021 ◽  
Vol 22 (14) ◽  
pp. 7565
Author(s):  
Kyungho Woo ◽  
Dong Ho Kim ◽  
Man Hwan Oh ◽  
Ho Sung Park ◽  
Chul Hee Choi
Keyword(s):  
Bone Marrow ◽  
Quorum Sensing ◽  
Deletion Mutant ◽  
Transmembrane Protein ◽  
Cell Communication ◽  
Homoserine Lactone ◽  
Host Responses ◽  
Wild Type ◽  
Mutant Strains

Quorum sensing of Acinetobacter nosocomialis for cell-to-cell communication produces N-3-hydroxy dodecanoyl-DL-homoserine lactone (OH-dDHL) by an AnoR/I two-component system. However, OH-dDHL-driven apoptotic mechanisms in hosts have not been clearly defined. Here, we investigated the induction of apoptosis signaling pathways in bone marrow-derived macrophages treated with synthetic OH-dDHL. Moreover, the quorum-sensing system for virulence regulation was evaluated in vivo using wild-type and anoI-deletion mutant strains. OH-dDHL decreased the viability of macrophage and epithelial cells in dose- and time-dependent manners. OH-dDHL induced Ca2+ efflux and caspase-12 activation by ER stress transmembrane protein (IRE1 and ATF6a p50) aggregation and induced mitochondrial dysfunction through reactive oxygen species (ROS) production, which caused cytochrome c to leak. Pretreatment with a pan-caspase inhibitor reduced caspase-3, -8, and -9, which were activated by OH-dDHL. Pro-inflammatory cytokine and paraoxonase-2 (PON2) gene expression were increased by OH-dDHL. We showed that the anoI-deletion mutant strains have less intracellular invasion compared to the wild-type strain, and their virulence, such as colonization and dissemination, was decreased in vivo. Consequently, these findings revealed that OH-dDHL, as a virulence factor, contributes to bacterial infection and survival as well as the modification of host responses in the early stages of infection.

scholarly journals Villidin, a Novel WD-repeat and Villin-related Protein fromDictyostelium,Is Associated with Membranes and the Cytoskeleton

2003 ◽  
Vol 14 (7) ◽  
pp. 2716-2727 ◽  
Author(s):  
Annika Gloss ◽  
Francisco Rivero ◽  
Nandkumar Khaire ◽  
Rolf Müller ◽  
William F. Loomis ◽  
...  
Keyword(s):  
Actin Binding ◽  
Similar Distribution ◽  
Fruiting Bodies ◽  
Targeted Deletion ◽  
C Terminus ◽  
Its Gene ◽  
Mutant Strains ◽  
Protein Functions ◽  
Wd Repeats ◽  
Actin Binding Site

Villidin is a novel multidomain protein (190 kDa) from Dictyostelium amoebae containing WD repeats at its N-terminus, three PH domains in the middle of the molecule, and five gelsolin-like segments at the C-terminus, followed by a villin-like headpiece. Villidin mRNA and protein are present in low amounts during growth and early aggregation, but increase during development and reach their highest levels at the tipped mound stage. The protein is present in the cytosol as well as in the cytoskeletal and membrane fractions. GFP-tagged full-length villidin exhibits a similar distribution as native villidin, including a distinct colocalization with Golgi structures. Interestingly, GFP fusions with the gelsolin/villin-like region are uniformly dispersed in the cytoplasm, whereas GFP fusions of the N-terminal WD repeats codistribute with F-actin and are associated with the Triton-insoluble cytoskeleton. Strains lacking villidin because of targeted deletion of its gene grow normally and can develop into fruiting bodies. However, cell motility is reduced during aggregation and phototaxis is impaired in the mutant strains. We conclude that villidin harbors a major F-actin binding site in the N-terminal domain and not in the villin-like region as expected; association of villidin with vesicular membranes suggests that the protein functions as a linker between membranes and the actin cytoskeleton.

scholarly journals β-Galactosidase from Termination and Deletion Mutant Strains

1974 ◽  
Vol 120 (1) ◽  
pp. 466-474 ◽  
Author(s):  
Merna R. Villarejo ◽  
Irving Zabin
Keyword(s):  
Deletion Mutant ◽  
Mutant Strains

A new phosphotriesterase from Sulfolobus acidocaldarius and its comparison with the homologue from Sulfolobus solfataricus

Biochimie ◽  
2007 ◽  
Vol 89 (5) ◽  
pp. 625-636 ◽  
Author(s):  
E PORZIO ◽  
L MERONE ◽  
L MANDRICH ◽  
M ROSSI ◽  
G MANCO
Keyword(s):  
Sulfolobus Solfataricus ◽  
Sulfolobus Acidocaldarius

scholarly journals The Impact of Pyroglutamate:Sulfolobus acidocaldariusHas a Growth Advantage overSaccharolobus solfataricusin Glutamate-Containing Media

Archaea ◽  
2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Anna M. Vetter ◽  
Julia Helmecke ◽  
Dietmar Schomburg ◽  
Meina Neumann-Schaal
Keyword(s):  
Carbon Source ◽  
Sole Carbon Source ◽  
Sulfolobus Solfataricus ◽  
Cell Extract ◽  
Sulfolobus Acidocaldarius ◽  
Intracellular Accumulation ◽  
Efficient Utilization ◽  
Crude Cell Extract ◽  
Toxic Metabolites ◽  
The Impact

Microorganisms are well adapted to their habitat but are partially sensitive to toxic metabolites or abiotic compounds secreted by other organisms or chemically formed under the respective environmental conditions. Thermoacidophiles are challenged by pyroglutamate, a lactam that is spontaneously formed by cyclization of glutamate under aerobic thermoacidophilic conditions. It is known that growth of the thermoacidophilic crenarchaeonSaccharolobus solfataricus(formerlySulfolobus solfataricus) is completely inhibited by pyroglutamate. In the present study, we investigated the effect of pyroglutamate on the growth ofS. solfataricusand the closely related crenarchaeonSulfolobus acidocaldarius.In contrast toS. solfataricus,S. acidocaldariuswas successfully cultivated with pyroglutamate as a sole carbon source. Bioinformatical analyses showed that both members of theSulfolobaceaehave at least one candidate for a 5-oxoprolinase, which catalyses the ATP-dependent conversion of pyroglutamate to glutamate. InS. solfataricus, we observed the intracellular accumulation of pyroglutamate and crude cell extract assays showed a less effective degradation of pyroglutamate. Apparently,S. acidocaldariusseems to be less versatile regarding carbohydrates and prefers peptidolytic growth compared toS. solfataricus. Concludingly,S. acidocaldariusexhibits a more efficient utilization of pyroglutamate and is not inhibited by this compound, making it a better candidate for applications with glutamate-containing media at high temperatures.

scholarly journals Nonsense suppression of the major rhodopsin gene of Drosophila.

Genetics ◽  
1992 ◽  
Vol 130 (3) ◽  
pp. 585-595 ◽  
Author(s):  
T Washburn ◽  
J E O'Tousa
Keyword(s):  
Deletion Mutant ◽  
Trna Gene ◽  
Site Directed Mutagenesis ◽  
P Element ◽  
Nonsense Suppression ◽  
Nonsense Mutations ◽  
Rhodopsin Gene ◽  
Translational Readthrough ◽  
Mutant Strains ◽  
Nonsense Suppressors

Abstract We placed UAA, UAG and UGA nonsense mutations at two leucine codons, Leu205 and Leu309, in Drosophila's major rhodopsin gene, ninaE, by site-directed mutagenesis, and then created the corresponding mutants by P element-mediated transformation of a ninaE deficiency strain. In the absence of a genetic suppressor, flies harboring any of the nonsense mutations at the 309 site, but not the 205 site, show increased rhodopsin activity. Additionally, all flies with nonsense mutations at either site have better rhabdomere structure than does the ninaE deficiency strain. Construction and analysis of a 3'-deletion mutant of ninaE indicates that translational readthrough accounts for the extra photoreceptor activity of the ninaE309 alleles and that truncated opsins are responsible for the improved rhabdomere structure. The presence of leucine-inserting tRNA nonsense suppressors DtLa Su+ and DtLb Su+ in the mutant strains produced a small increase (less than 0.04%) in functional rhodopsin. The opal (UGA) suppressor derived from the DtLa tRNA gene is more efficient than the amber (UAG) or opal suppressor derived from the DtLb gene, and both DtLa and DtLb derived suppressors are more efficient at site 205 than 309.

scholarly journals Transcriptome Analysis of Aspergillus nidulans Exposed to Camptothecin-Induced DNA Damage

2006 ◽  
Vol 5 (10) ◽  
pp. 1688-1704 ◽  
Author(s):  
Iran Malavazi ◽  
Marcela Savoldi ◽  
Sônia Marli Zingaretti Di Mauro ◽  
Carlos Frederico Martins Menck ◽  
Steven D. Harris ◽  
...  
Keyword(s):  
Dna Damage ◽  
Aspergillus Nidulans ◽  
Mutant Strain ◽  
Deletion Mutant ◽  
Time Course ◽  
Excision Repair ◽  
Wild Type ◽  
Significant Difference ◽  
Mutant Strains ◽  
In The Wild

ABSTRACT We have used an Aspergillus nidulans macroarray carrying sequences of 2,787 genes from this fungus to monitor gene expression of both wild-type and uvsB ATR (the homologue of the ATR gene) deletion mutant strains in a time course exposure to camptothecin (CPT). The results revealed a total of 1,512 and 1,700 genes in the wild-type and uvsB ATR deletion mutant strains that displayed a statistically significant difference at at least one experimental time point. We characterized six genes that have increased mRNA expression in the presence of CPT in the wild-type strain relative to the uvsB ATR mutant strain: fhdA (encoding a forkhead-associated domain protein), tprA (encoding a hypothetical protein that contains a tetratrico peptide repeat), mshA (encoding a MutS homologue involved in mismatch repair), phbA (encoding a prohibitin homologue), uvsC RAD51 (the homologue of the RAD51 gene), and cshA (encoding a homologue of the excision repair protein ERCC-6 [Cockayne's syndrome protein]). The induced transcript levels of these genes in the presence of CPT require uvsB ATR. These genes were deleted, and surprisingly, only the ΔuvsC mutant strain was sensitive to CPT; however, the others displayed sensitivity to a range of DNA-damaging and oxidative stress agents. These results indicate that the selected genes when inactivated display very complex and heterogeneous sensitivity behavior during growth in the presence of agents that directly or indirectly cause DNA damage. Moreover, with the exception of UvsC, deletion of each of these genes partially suppressed the sensitivity of the ΔuvsB strain to menadione and paraquat. Our results provide the first insight into the overall complexity of the response to DNA damage in filamentous fungi and suggest that multiple pathways may act in parallel to mediate DNA repair.

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