HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 49, 49113-49118, December 5, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/49/49113    most recent M308652200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chen, Q. Articles by Haddad, G. G. Search for Related Content PubMed PubMed Citation Articles by Chen, Q. Articles by Haddad, G. G. Social Bookmarking                      What's this?

Expression of Drosophila Trehalose-Phosphate Synthase in HEK-293 Cells Increases Hypoxia Tolerance^*

Qiaofang Chen, Kevin L. Behar, Tian Xu¶, Chenhao Fan, and Gabriel G. Haddad||**

From the Departments of Pediatrics and ^||Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461 and the Department of Psychiatry and ^¶Howard Hughes Medical Institute, Department of Genetics and Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06520

Increasing hypoxia tolerance in mammalian cells is potentially of major importance, but it has not been feasible thus far. The disaccharide trehalose, which accumulates dramatically during heat shock, enhances thermotolerance and reduces aggregation of denatured proteins. Previous studies from our laboratory showed that over-expression of Drosophila trehalose-phosphate synthase (dtps1) increases the trehalose level and anoxia tolerance in flies. To determine whether trehalose can protect against anoxic injury in mammalian cells, we transfected the dtps1 gene into human HEK-293 cells using the recombinant plasmid pcDNA3.1(–)-dtps1 and obtained more than 20 stable cell strains. Glucose starvation in culture showed that HEK-293 cells transfected with pcDNA3.1(–)-dtps1 (HEK-dtps1) do not metabolize intracellular trehalose, and, interestingly, these cells accumulated intracellular trehalose during hypoxic exposure. In contrast to HEK-293 cells transfected with pcDNA3.1(–) (HEK-v), cells with trehalose were more resistant to low oxygen stress (1% O₂). To elucidate how trehalose protects cells from anoxic injury, we assayed protein solubility and the amount of ubiquitinated proteins. There was three times more insoluble protein in HEK-v than in HEK-dtps1 after 3 days of exposure to low O₂. The amount of Na⁺-K⁺ ATPase present in the insoluble proteins dramatically increased in HEK-v cells after 2 and 3 days of exposure, whereas there was no significant change in HEK-dtps1 cells. Ubiquitinated proteins increased dramatically in HEK-v cells after 2 and 3 days of exposure but not in HEK-dtps1 cells over the same period. Our results indicate that increased trehalose in mammalian cells following transfection by the Drosophila tps1 gene protects cells from hypoxic injury. The mechanism of this protection is likely related to a decrease in protein denaturation, through protein-trehalose interactions, resulting in enhanced cellular recovery from hypoxic stress.

Received for publication, August 6, 2003 , and in revised form, September 8, 2003.

^* This work was supported by National Institutes of Health Grants P01-HD32573-09 and R01-NS37754 (to G. G. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^** To whom correspondence should be addressed: Depts. of Pediatrics and Neuroscience, Albert Einstein College of Medicine, 1410 Pelham Pkwy., Bronx, NY 10461. Tel.: 718-430-8094; Fax: 718-430-2199; E-mail: Ghaddad{at}aecom.yu.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. D. Higgins, E. Bancalari, M. Willinger, and T. N.K. Raju Executive Summary of the Workshop on Oxygen in Neonatal Therapies: Controversies and Opportunities for Research Pediatrics, April 1, 2007; 119(4): 790 - 796. [Abstract] [Full Text] [PDF]

				S. Sarkar, J. E. Davies, Z. Huang, A. Tunnacliffe, and D. C. Rubinsztein Trehalose, a Novel mTOR-independent Autophagy Enhancer, Accelerates the Clearance of Mutant Huntingtin and {alpha}-Synuclein J. Biol. Chem., February 23, 2007; 282(8): 5641 - 5652. [Abstract] [Full Text] [PDF]

				G. Liu, J. Roy, and E. A. Johnson Identification and function of hypoxia-response genes in Drosophila melanogaster Physiol Genomics, March 13, 2006; 25(1): 134 - 141. [Abstract] [Full Text] [PDF]

				G. G. Haddad Tolerance to low O2: lessons from invertebrate genetic models Exp Physiol, March 1, 2006; 91(2): 277 - 282. [Abstract] [Full Text] [PDF]

				S. T. Lamitina and K. Strange Transcriptional targets of DAF-16 insulin signaling pathway protect C. elegans from extreme hypertonic stress Am J Physiol Cell Physiol, February 1, 2005; 288(2): C467 - C474. [Abstract] [Full Text] [PDF]

				Q. Chen and G. G. Haddad Role of trehalose phosphate synthase and trehalose during hypoxia: from flies to mammals J. Exp. Biol., August 15, 2004; 207(18): 3125 - 3129. [Abstract] [Full Text] [PDF]