Reconstruction of central carbon metabolism in Sulfolobus solfataricus using a two-dimensional gel electrophoresis map, stable isotope labelling and DNA microarray analysis

In the last decade, an increasing number of sequenced archaeal genomes have become available, opening up the possibility for functional genomic analyses. Here, we reconstructed the central carbon metabolism in the hyperthermophilic crenarchaeon Sulfolobus solfataricus (glycolysis, gluconeogenesis an...

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Bibliographic Details
Published in:	Proteomics (Weinheim) Vol. 6; no. 5; pp. 1518 - 1529
Main Authors:	Snijders, Ambrosius P. L., Walther, Jasper, Peter, Stefan, Kinnman, Iris, de Vos, Marjon G. J., van de Werken, Harmen J. G., Brouns, Stan J. J., van der Oost, John, Wright, Phillip C.
Format:	Journal Article
Language:	English
Published:	Weinheim WILEY-VCH Verlag 01-03-2006 WILEY‐VCH Verlag Wiley-VCH
Subjects:	Analytical, structural and metabolic biochemistry Archaea Archaeal Proteins - chemistry Archaeal Proteins - genetics Archaeal Proteins - metabolism archaebacterium sulfolobus Biological and medical sciences Carbon - metabolism citrate synthase citric-acid cycle DNA microarray Electrophoresis, Gel, Two-Dimensional entner-doudoroff pathway Fundamental and applied biological sciences. Psychology Gene Expression Profiling Glycolysis glycolytic pathways hyperthermophilic archaea Isotopes - chemistry Mass Spectrometry Metabolic labelling Miscellaneous Molecular Sequence Data Oligonucleotide Array Sequence Analysis phosphoenolpyruvate carboxylase Proteins Sulfolobus solfataricus Sulfolobus solfataricus - chemistry Sulfolobus solfataricus - genetics Sulfolobus solfataricus - metabolism thermoacidophilic crenarchaeon thermoplasma-acidophilum thermoproteus-tenax Isotope labelling Archaeobacteria Archaea DNA chip Sulfolobus solfataricus Metabolism Sulfolobaceae microarray / Glycolysis / Mass spectrometry / Metabolic labelling Proteomics Glycolysis Bacteria Sulfolobales Two dimensional electrophoresis Mass spectrometry
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Summary:	In the last decade, an increasing number of sequenced archaeal genomes have become available, opening up the possibility for functional genomic analyses. Here, we reconstructed the central carbon metabolism in the hyperthermophilic crenarchaeon Sulfolobus solfataricus (glycolysis, gluconeogenesis and tricarboxylic acid cycle) on the basis of genomic, proteomic, transcriptomic and biochemical data. A 2‐DE reference map of S. solfataricus grown on glucose, consisting of 325 unique ORFs in 255 protein spots, was created to facilitate this study. The map was then used for a differential expression study based on 15N metabolic labelling (yeast extract + tryptone‐grown cells (YT) vs. glucose‐grown cells (G)). In addition, the expression ratio of the genes involved in carbon metabolism was studied using DNA microarrays. Surprisingly, only 3 and 14% of the genes and proteins, respectively, involved in central carbon metabolism showed a greater than two‐fold change in expression level. All results are discussed in the light of the current understanding of central carbon metabolism in S. solfataricus and will help to obtain a system‐wide understanding of this organism.
Bibliography:	istex:75A4C023CE6E3CC369154838AB13360CC7332447 ark:/67375/WNG-8VJ1HJ2V-P ArticleID:PMIC200402070 Both authors contributed equally to this work. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1615-9853 1615-9861
DOI:	10.1002/pmic.200402070