Disparity between changes in mRNA abundance and enzyme activity in Corynebacterium glutamicum: implications for DNA microarray analysis (original) (raw)

Abstract

The relationship between changes in mRNA abundance and enzyme activity was determined for three genes over a span of nearly 3 h during amino acid production in Corynebacterium glutamicum. Gene expression changes during C. glutamicum fermentations were examined by complementary DNA (cDNA) microarrays and by a second method for quantitating RNA levels, competitive reverse transcriptase-PCR (RT-PCR). The results obtained independently by both methods were compared and found to be in agreement, thus validating the quantitative potential of DNA microarrays for gene expression profiling. Evidence of a disparity between mRNA abundance and enzyme activity is presented and supports our belief that it is difficult to generally predict protein activity from quantitative transcriptome data. Homoserine dehydrogenase, threonine dehydratase, and homoserine kinase are enzymes involved in the biosynthesis of l-isoleucine and other aspartate-derived amino acids in C. glutamicum. Our data suggest that different underlying regulatory mechanisms may be connected with the expression of the genes encoding each of these three enzymes. Indeed, whereas in one case the increases in enzyme activity exceeded those in the corresponding mRNA abundance, in another case large increases in the levels of gene expression were not congruent with changes in enzyme activity.

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Acknowledgements

We gratefully acknowledge Sean Milton of the BioMicro Center at MIT for his technical expertise. The experiments presented in this paper comply with the current laws of the country in which they were performed.

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Author notes

1. C. Glanemann
   Present address: Institute for Genetics, University of Cologne, Zülpicherstrasse 47, 50674 , Cologne, Germany

Authors and Affiliations

1. Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA
   C. Glanemann, A. Loos, N. Gorret, L. B. Willis, X. M. O'Brien, P. A. Lessard & A. J. Sinskey
2. BioMicro Center, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA
   A. Loos

Authors

1. C. Glanemann
2. A. Loos
3. N. Gorret
4. L. B. Willis
5. X. M. O'Brien
6. P. A. Lessard
7. A. J. Sinskey

Corresponding author

Correspondence toA. J. Sinskey.

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Glanemann, C., Loos, A., Gorret, N. et al. Disparity between changes in mRNA abundance and enzyme activity in Corynebacterium glutamicum:implications for DNA microarray analysis.Appl Microbiol Biotechnol 61, 61–68 (2003). https://doi.org/10.1007/s00253-002-1191-5

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• Received: 08 August 2002
• Revised: 25 October 2002
• Accepted: 08 November 2002
• Published: 21 December 2002
• Issue date: March 2003
• DOI: https://doi.org/10.1007/s00253-002-1191-5

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