Transcriptome analysis of acetate metabolism in Corynebacterium glutamicum using a newly developed metabolic array.
Following the determination of the whole-genome sequence of Corynebacterium glutamicum, we have developed a DNA array to extensively investigate gene expression and regulation relevant to carbon metabolism. For this purpose, a total of 120 C. glutamicum genes, including those in central metabolism and amino acid biosyntheses, were amplified by PCR and printed onto glass slides. The resulting array, designated a "metabolic array", was used for hybridization with fluorescently labeled cDNA probes generated by reverse transcription from total RNA samples. As the first demonstration of transcriptome analysis in this industrially important microorganism, we applied the metabolic array to study differential transcription profiles between cells grown on glucose and on acetate as the sole carbon source. The changes in gene expression observed for the known acetate-regulated genes (aceA, aceB, pta, and ack) were well consistent with the literature data of northern analyses and enzyme assays, indicating the utility of the metabolic array in transcriptome analysis of C. glutamicum. In addition to the known responses, many previously unrecognized co-regulated genes were identified. For example, several TCA cycle genes, such as gltA, sdhA, sdhB, fumH, and mdh, and the gluconeogenic gene pck were up-regulated in the acetate medium. On the other hand, a few genes involved in glycolysis and the pentose phosphate pathway, as well as many amino acid biosynthetic genes, were down-regulated in acetate. Furthermore, two gap genes, gapA and gapB, were found to be inversely regulated, suggesting the presence of a new regulatory step for carbon metabolism between glycolysis and gluconeogenesis.
Hayashi
Mikiro
M
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co, Ltd, Machida, Japan.
Mizoguchi
Hiroshi
H
Shiraishi
Norihiko
N
Obayashi
Masaya
M
Nakagawa
Satoshi
S
Imai
Jun-ichi
J
Watanabe
Shinya
S
Ota
Toshio
T
Ikeda
Masato
M
D000085:Acetates / Q000378:metabolism*; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000235:genetics* / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D005947:Glucose / Q000378:metabolism; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics*
D000085:Acetates; D018076:DNA, Complementary; D012329:RNA, Bacterial; D005947:Glucose