| An AMP nucleosidase gene knockout in Escherichia coli elevates intracellular ATP levels and increases cold tolerance. | |
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MedLine Citation:
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PMID: 18029299 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Disparate psychrophiles (e.g. glacier ice worms, bacteria, algae and fungi) elevate steady-state intracellular ATP levels as temperatures decline, which has been interpreted as a compensatory mechanism to offset reductions in molecular motion and Gibb's free energy of ATP hydrolysis. In this study, we sought to manipulate steady-state ATP levels in the mesophilic bacterium, Escherichia coli, to investigate the relationship between cold temperature survivability and elevated intracellular ATP. Based on known energetic pathways and feedback loops, we targeted the AMP nucleotidase (amn) gene, which is thought to encode the primary AMP degradative enzyme in prokaryotes. By knocking out amn in wild-type E. coli DY330 cells using recombineering methodology, we generated a mutant (AMNk) that elevated intracellular ATP levels by more than 30% across its viable temperature range. As temperature was lowered, the relative ATP disparity between AMNk and DY330 cells increased to approximately 66% at 10 degrees C, and was approximately 100% after storage at 0 degrees C for 5-7 days. AMNk cells stored at 0 degrees C for 7 days displayed approximately fivefold higher cell viability than wild-type DY330 cells treated in the same manner. |
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Authors:
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Brittany A Morrison; Daniel H Shain |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Biology letters Volume: 4 ISSN: 1744-9561 ISO Abbreviation: Biol. Lett. Publication Date: 2008 Feb |
Date Detail:
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Created Date: 2008-01-09 Completed Date: 2008-03-21 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 101247722 Medline TA: Biol Lett Country: England |
Other Details:
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Languages: eng Pagination: 53-6 Citation Subset: IM |
Affiliation:
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Biology Department, Rutgers The State University of New Jersey, Camden, NJ 08102, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adaptation, Physiological
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genetics Adenosine Triphosphate / metabolism* Cold Temperature* Escherichia coli / enzymology*, genetics* Gene Deletion Gene Expression Regulation, Bacterial N-Glycosyl Hydrolases / genetics* |
| Chemical | |
Reg. No./Substance:
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56-65-5/Adenosine Triphosphate; EC 3.2.2.-/N-Glycosyl Hydrolases; EC 3.2.2.4/AMP nucleosidase |
| Comments/Corrections | |
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