TY - JOUR
T1 - Reconstruction of central carbon metabolism in Sulfolobus solfataricus using a two-dimensional gel electrophoresis map, stable isotope labelling and DNA microarray analysis
AU - Snijders, B.P.L.
AU - Walther, J.
AU - Peter, S.
AU - Kinnman, I.
AU - de Vos, M.J.G.
AU - van de Werken, H.J.G.
AU - Brouns, S.J.J.
AU - van der Oost, J.
AU - Wright, P.C.
PY - 2006
Y1 - 2006
N2 - 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 (15)N 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.
AB - 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 (15)N 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.
KW - entner-doudoroff pathway
KW - citric-acid cycle
KW - hyperthermophilic archaea
KW - archaebacterium sulfolobus
KW - citrate synthase
KW - phosphoenolpyruvate carboxylase
KW - thermoacidophilic crenarchaeon
KW - thermoplasma-acidophilum
KW - thermoproteus-tenax
KW - glycolytic pathways
U2 - 10.1002/pmic.200402070
DO - 10.1002/pmic.200402070
M3 - Article
SN - 1615-9853
VL - 6
SP - 1518
EP - 1529
JO - Proteomics
JF - Proteomics
IS - 5
ER -