Chhabra Swapnil R, Shockley Keith R, Conners Shannon B, Scott Kevin L, Wolfinger Russell D, Kelly Robert M
Department of Chemical Engineering, North Carolina State University, Raleigh, North Carolina 27695, USA.
J Biol Chem. 2003 Feb 28;278(9):7540-52. doi: 10.1074/jbc.M211748200. Epub 2002 Dec 9.
The hyperthermophilic bacterium Thermotoga maritima MSB8 was grown on a variety of carbohydrates to determine the influence of carbon and energy source on differential gene expression. Despite the fact that T. maritima has been phylogenetically characterized as a primitive microorganism from an evolutionary perspective, results here suggest that it has versatile and discriminating mechanisms for regulating and effecting complex carbohydrate utilization. Growth of T. maritima on monosaccharides was found to be slower than growth on polysaccharides, although growth to cell densities of 10(8) to 10(9) cells/ml was observed on all carbohydrates tested. Differential expression of genes encoding carbohydrate-active proteins encoded in the T. maritima genome was followed using a targeted cDNA microarray in conjunction with mixed model statistical analysis. Coordinated regulation of genes responding to specific carbohydrates was noted. Although glucose generally repressed expression of all glycoside hydrolase genes, other sugars induced or repressed these genes to varying extents. Expression profiles of most endo-acting glycoside hydrolase genes correlated well with their reported biochemical properties, although exo-acting glycoside hydrolase genes displayed less specific expression patterns. Genes encoding selected putative ABC sugar transporters were found to respond to specific carbohydrates, and in some cases putative oligopeptide transporter genes were also found to respond to specific sugar substrates. Several genes encoding putative transcriptional regulators were expressed during growth on specific sugars, thus suggesting functional assignments. The transcriptional response of T. maritima to specific carbohydrate growth substrates indicated that sugar backbone- and linkage-specific regulatory networks are operational in this organism during the uptake and utilization of carbohydrate substrates. Furthermore, the wide ranging collection of such networks in T. maritima suggests that this organism is capable of adapting to a variety of growth environments containing carbohydrate growth substrates.
嗜热栖热菌(Thermotoga maritima)MSB8在多种碳水化合物上生长,以确定碳源和能源对差异基因表达的影响。尽管从进化角度来看,嗜热栖热菌在系统发育上被归类为原始微生物,但此处的结果表明,它具有多种且有区别的机制来调节和实现复杂碳水化合物的利用。发现嗜热栖热菌在单糖上的生长比在多糖上慢,不过在所有测试的碳水化合物上都观察到其生长至细胞密度达到10⁸至10⁹个细胞/毫升。使用靶向cDNA微阵列结合混合模型统计分析,追踪嗜热栖热菌基因组中编码碳水化合物活性蛋白的基因的差异表达。注意到对特定碳水化合物有反应的基因的协调调节。虽然葡萄糖通常会抑制所有糖苷水解酶基因的表达,但其他糖类会不同程度地诱导或抑制这些基因。大多数内切糖苷水解酶基因的表达谱与其报道的生化特性相关性良好,尽管外切糖苷水解酶基因显示出不太特异的表达模式。发现编码选定推定ABC糖转运蛋白的基因对特定碳水化合物有反应,在某些情况下还发现推定的寡肽转运蛋白基因对特定糖底物有反应。在特定糖类上生长期间,几个编码推定转录调节因子的基因被表达,因此表明了功能归属。嗜热栖热菌对特定碳水化合物生长底物的转录反应表明,在该生物体摄取和利用碳水化合物底物期间,糖主链和连接特异性调节网络在起作用。此外,嗜热栖热菌中此类网络的广泛收集表明,该生物体能够适应含有碳水化合物生长底物的多种生长环境。