Hovey Raymond, Lentes Sabine, Ehrenreich Armin, Salmon Kirsty, Saba Karla, Gottschalk Gerhard, Gunsalus Robert P, Deppenmeier Uwe
Department of Biological Sciences, University of Wisconsin-Milwaukee, N. Maryland Ave 3209, Milwaukee, WI 53211, USA.
Mol Genet Genomics. 2005 May;273(3):225-39. doi: 10.1007/s00438-005-1126-9. Epub 2005 Apr 7.
Methansarcina mazei Gö1 DNA arrays were constructed and used to evaluate the genomic expression patterns of cells grown on either of two alternative methanogenic substrates, acetate or methanol, as sole carbon and energy source. Analysis of differential transcription across the genome revealed two functionally grouped sets of genes that parallel the central biochemical pathways in, and reflect many known features of, acetate and methanol metabolism. These include the acetate-induced genes encoding acetate activating enzymes, acetyl-CoA synthase/CO dehydrogenase, and carbonic anhydrase. Interestingly, additional genes expressed at significantly higher levels during growth on acetate included two energy-conserving complexes (the Ech hydrogenase, and the A1A0-type ATP synthase). Many previously unknown features included the induction by acetate of genes coding for ferredoxins and flavoproteins, an aldehyde:ferredoxin oxidoreductase, enzymes for the synthesis of aromatic amino acids, and components of iron, cobalt and oligopeptide uptake systems. In contrast, methanol-grown cells exhibited elevated expression of genes assigned to the methylotrophic pathway of methanogenesis. Expression of genes for components of the translation apparatus was also elevated in cells grown in the methanol medium relative to acetate, and was correlated with the faster growth rate observed on the former substrate. These experiments provide the first comprehensive insight into substrate-dependent gene expression in a methanogenic archaeon. This genome-wide approach, coupled with the complementary molecular and biochemical tools, should greatly accelerate the exploration of Methanosarcina cell physiology, given the present modest level of our knowledge of these large archaeal genomes.
构建了马氏甲烷八叠球菌Gö1 DNA阵列,并用于评估以两种替代性产甲烷底物(乙酸盐或甲醇)中的任何一种作为唯一碳源和能源生长的细胞的基因组表达模式。对全基因组差异转录的分析揭示了两组功能分组的基因,它们与乙酸盐和甲醇代谢中的中心生化途径平行,并反映了其中许多已知特征。其中包括乙酸盐诱导的基因,这些基因编码乙酸盐激活酶、乙酰辅酶A合酶/一氧化碳脱氢酶和碳酸酐酶。有趣的是,在乙酸盐上生长期间表达水平显著更高的其他基因包括两种能量保守复合物(Ech氢化酶和A1A0型ATP合酶)。许多以前未知的特征包括乙酸盐对编码铁氧化还原蛋白和黄素蛋白、醛:铁氧化还原蛋白氧化还原酶、芳香族氨基酸合成酶以及铁、钴和寡肽摄取系统成分的基因的诱导。相比之下,以甲醇生长的细胞表现出参与甲烷生成甲基营养途径的基因表达升高。相对于乙酸盐,在甲醇培养基中生长的细胞中翻译装置成分的基因表达也升高,并且与在前一种底物上观察到的更快生长速率相关。这些实验首次全面深入了解了产甲烷古菌中底物依赖性基因表达。鉴于我们目前对这些大型古菌基因组的了解程度有限,这种全基因组方法与互补的分子和生化工具相结合,应该会大大加速对甲烷八叠球菌细胞生理学的探索。