Adenosine thiamine triphosphate accumulates in Escherichia coli cells in response to specific conditions of metabolic stress.

Abstract

BACKGROUND:

E. coli cells are rich in thiamine, most of it in the form of the cofactor thiamine diphosphate (ThDP). Free ThDP is the precursor for two triphosphorylated derivatives, thiamine triphosphate (ThTP) and the newly discovered adenosine thiamine triphosphate (AThTP). While, ThTP accumulation requires oxidation of a carbon source, AThTP slowly accumulates in response to carbon starvation, reaching approximately 15% of total thiamine. Here, we address the question whether AThTP accumulation in E. coli is triggered by the absence of a carbon source in the medium, the resulting drop in energy charge or other forms of metabolic stress.

RESULTS:

In minimal M9 medium, E. coli cells produce AThTP not only when energy substrates are lacking but also when their metabolization is inhibited. Thus AThTP accumulates in the presence of glucose, when glycolysis is blocked by iodoacetate, or in the presence lactate, when respiration is blocked by cyanide or anoxia. In both cases, ATP synthesis is impaired, but AThTP accumulation does not appear to be a direct consequence of reduced ATP levels. Indeed, in the CV2 E. coli strain (containing a thermolabile adenylate kinase), the ATP content is very low at 37 degrees C, even in the presence of metabolizable substrates (glucose or lactate) and under these conditions, the cells produce ThTP but not AThTP. Furthermore, we show that ThTP inhibits AThTP accumulation. Therefore, we conclude that a low energy charge is not sufficient to trigger AThTP accumulation and the latter can only accumulate under conditions where no ThTP is synthesized. We further show that AThTP production can also be induced by the uncoupler CCCP but, unexpectedly, this requires the presence of pyruvate or a substrate yielding pyruvate (such a D-glucose or L-lactate). Under the conditions described, AThTP production is not different when RelA or SpoT mutants are used.

CONCLUSIONS:

In E. coli, AThTP accumulates in response to two different conditions of metabolic stress: lack of energy substrates (or inhibition of their metabolization) and uncoupled pyruvate oxidation. Both conditions prevent bacterial growth. There is no obvious link with the stringent response or catabolite repression.

• Authors: Tiziana Gigliobianco, Bernard Lakaye, Pierre Wins, Benaïssa El Moualij, Willy Zorzi, Lucien Bettendorff
• Journal: BMC microbiology (BMC Microbiol) Vol. 10 Pg. 148 (May 21 2010) ISSN: 1471-2180 [Electronic] England
• PMID: 20492686 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Culture Media
• Uncoupling Agents
• adenosine thiamine triphosphate
• Lactic Acid
• Thiamine Triphosphate
• Carbonyl Cyanide m-Chlorophenyl Hydrazone
• Carbon
• Pyruvic Acid
• Adenosine Triphosphate
• Glucose

Topics

• Adenosine Triphosphate (biosynthesis, metabolism)
• Carbon (metabolism)
• Carbonyl Cyanide m-Chlorophenyl Hydrazone (pharmacology)
• Culture Media (chemistry)
• Energy Metabolism
• Escherichia coli (growth & development, metabolism, physiology)
• Glucose (metabolism)
• Lactic Acid (metabolism)
• Pyruvic Acid (metabolism)
• Stress, Physiological
• Thiamine Triphosphate (metabolism)
• Uncoupling Agents (pharmacology)