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Hodgskiss, L. H.

Publications and source records attributed to Hodgskiss, L. H..

4 recordsLinked to original sources

Metabolic Response of a Chemolithoautotrophic Archaeon to Carbon Limitation

The ubiquitously distributed ammonia-oxidizing archaea generate energy from ammonia and build cell mass from inorganic carbon sources, thereby contributing to both the global nitrogen and carbon cycles. However, little is known about the regulation of their predicted core carbon metabolism. A thermodynamic model for Nitrososphaera viennensis was developed to estimate the consumption of inorganic carbon in relation to ammonia consumed for energy and was tested experimentally by growing cells in carbon-limited and excess conditions. A combined proteomic and metabolomic approach of the experimental conditions revealed distinct metabolic adaptation depending on the amount of carbon supplied, either in a catalase or pyruvate background as a reactive oxygen species scavenger. Integration of protein and metabolite dynamics revealed a cellular strategy under carbon limitation to maintain a pool of amino acids and an upregulation of proteins necessary for translation initiation to stay primed for protein synthesis. The combination of modelling and functional genomics fills gaps in the understanding of the central metabolism and its regulation in a chemolithoautotrophic, ammonia-oxidizing archaeon even in the absence of available genetic tools.

microbiology↗

Biofilm lifestyle as a common trait of ammonia-oxidizing archaea

Although widespread in nature, growth in biofilms has been relatively little explored in the globally distributed ammonia oxidizing archaea (AOA). Here we investigated six representatives of three different terrestrial and marine clades of AOA in a longitudinal and quantitative study for their ability to form biofilm and studied gene expression patterns of three representatives. While all strains grew on a solid surface, soil strains exhibited the highest capacity for biofilm formation. Based on microscopic and gene expression data, two different colonization strategies could be distinguished. S-layer containing AOA (from both soil and marine habitats) initialized attachment as single cells and subsequently formed denser layers and three-dimensional structures, while the S-layer free species of the Nitrosocosmicus clade attached as suspended aggregates to the surface and henceforth showed fastest establishment of biofilm. Transcription profiles were significantly different between planktonic and biofilm growth in all strains and revealed individual reactions, often fulfilling shared functions. In particular the strong expression of different types of multicopper oxidases was observed in all strains indicating modifications of their cell coats. S-layer carrying AOA each additionally expressed a set of adhesion proteins supporting attachment. Detoxification of nitrous compounds, copper acquisition as well as the expression of transcription factor B were also shared reactions among biofilm producing strains. However, the majority of differentially expressed protein families was distinct among the three strains illustrating that individual solutions have evolved for the shared growth mode of biofilm formation in AOA, probably driven by the different ecological niches.

microbiology↗

The type III-B CRISPR-Cas System Affects Energy Metabolism and Adaptation in the Archaeon Saccharolobus solfataricus

Type III CRISPR-Cas immune systems that recognize and cleave extrachromosomal RNA when active, are particularly widespread in archaea. Mechanistically, these systems have the potential to regulate gene expression of host genes on a post-transcriptional level, but very little is known about any potential accessory roles of type III-B systems beyond immunity. We have created knockout mutants of a type III-B CRISPR-Cas complex in the thermoacidophilic archaeon Saccharolobus solfataricus to investigate potential secondary functions of the type III-B system. Deletion mutants exhibited an accelerate growth but were less quickly adaptable to changes in carbon sources in their growth media. In line with this phenotype, upregulated genes were significantly enriched in functional categories of energy production and conversion, as well as with carbohydrate or amino acid transport and metabolism in RNAseq studies. Generally, a significant accumulation of genes encoding transmembrane proteins in the upregulated proportion of the transcriptome suggests interconnections between the type III-B CRISPR-Cas system and various membrane-associated processes. Notably, the deletion mutants did not lose their general virus- or plasmid defense activities indicating that this particular system might have been partially adopted for cellular regulatory roles.

microbiology↗

Unexpected Complexity of the Ammonia Monooxygenase in Archaea

Ammonia oxidation as the first step of nitrification constitutes a critical process in the global nitrogen cycle. However, fundamental knowledge of its key enzyme, the copper-dependent ammonia monooxygenase is lacking, in particular for the environmentally abundant ammonia oxidizing archaea (AOA). Here, the structure of the enzyme is investigated by blue-native gel electrophoresis and proteomics from native membrane complexes of two AOA. Beside the known AmoABC subunits and the earlier predicted AmoX, two new protein subunits, AmoY and AmoZ, were identified. They are unique to AOA, highly conserved and co-regulated, and their genes are linked to other AMO subunit genes in streamlined AOA genomes. Modelling and in gel cross-link approaches support an overall protomer structure similar to the distantly related bacterial particulate methane monooxygenase indicating that AmoY and AmoZ serve an important structural and functional role. These data open avenues for further structure-function studies of this ecologically important key nitrification complex.

microbiology↗