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Specificity of amino acid regulated gene expression: analysis of genes subjected to either complete or single amino acid deprivation

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Abstract

Amino acid deprivation activates the amino acid response (AAR) pathway that enhances transcription of genes containing an amino acid response element (AARE). The present data reveal a quantitative difference in the response to deprivation of individual amino acids. The AAR leads to increased eukaryotic initiation factor 2α (eIF2α) phosphorylation and ATF4 translation. When HepG2 cells were deprived of an individual essential amino acid, p-eIF2α and activating transcription factor 4 were increased, but the correlation was relatively weak. Complete amino acid starvation in either Earle’s balanced salt solution or Krebs–Ringer bicarbonate buffer (KRB) resulted in activation of transcription driven by a SNAT2 genomic fragment that contained an AARE. However, for the KRB, a proportion of the transcription was AARE-independent suggesting that amino acid-independent mechanisms were responsible. Therefore, activation of AARE-driven transcription is triggered by a deficiency in any one of the essential amino acids, but the response is not uniform. Furthermore, caution must be exercised when using a medium completely devoid of amino acids.

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Abbreviations

AAR:

Amino acid response

AARE:

Amino acid response element

ASNS:

Asparagine synthetase

ATF3:

Activating transcription factor 3

ATF4:

Activating transcription factor 4

C/EBPβ:

CCAAT/enhancer-binding protein beta

CAT-1:

Cationic amino acid transporter

CHOP:

C/EBP homology protein

EAA:

Essential amino acids

EBSS:

Earle’s balanced salt solution

eIF2:

Eukaryotic initiation factor 2

GCN2:

General control non-derepressible

KRB:

Krebs–Ringer bicarbonate buffer

MEM:

Minimal essential medium

qPCR:

Quantitative real-time PCR

RT-PCR:

Reverse transcriptase-polymerase chain reaction

SNAT2:

System A sodium-dependent neutral amino acid transporter 2

VEGF:

Vascular endothelial growth factor

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Acknowledgments

This research was supported by grants to M.S. Kilberg from the Institute of Diabetes, Digestive and Kidney Diseases, the National Institutes of Health (DK-70647 and DK-52064). The authors wish to thank the other members of the laboratory for technical advice and helpful discussion.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Genetics Institute, Shands Cancer Center and Center for Nutritional Sciences, University of Florida, College of Medicine, Box 100245, Gainesville, FL, 32610-0245, USA

    S. S. Palii, C. E. Kays & M. S. Kilberg

  2. Unité de Nutrition Humaine, équipe Genes-Nutriments, INRA de Clermont-Theix, 63122, Saint Genès Champanelle, France

    C. Deval, A. Bruhat & P. Fafournoux

Authors
  1. S. S. Palii
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  2. C. E. Kays
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  3. C. Deval
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  4. A. Bruhat
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  5. P. Fafournoux
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  6. M. S. Kilberg
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Corresponding author

Correspondence to M. S. Kilberg.

Additional information

S. S. Palii and C. E. Kays have contributed equally to these studies.

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Palii, S.S., Kays, C.E., Deval, C. et al. Specificity of amino acid regulated gene expression: analysis of genes subjected to either complete or single amino acid deprivation. Amino Acids 37, 79–88 (2009). https://doi.org/10.1007/s00726-008-0199-2

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  • DOI: https://doi.org/10.1007/s00726-008-0199-2

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