Universität zu Köln

Pfeiffer, Carolin, Bach, Markus, Bauer, Tim, da Ponte, Julia Campos, Schoemig, Edgar and Gruendemann, Dirk (2015). Knockout of the ergothioneine transporter ETT in zebrafish results in increased 8-oxoguanine levels. Free Radic. Biol. Med., 83. S. 178 - 186. NEW YORK: ELSEVIER SCIENCE INC. ISSN 1873-4596

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Abstract

Ergothioneine (ET) is a natural compound that humans and other vertebrates must absorb from dietary sources. In general, ET is considered an intracellular antioxidant. However, the precise physiological purpose of ET and the consequences of ET deficiency are still unclear. The ergothioneine transporter ETT (human gene symbol SLC22A4) is a highly specific transporter for the uptake of ET. Here, we sought to identify and knock out ETT from zebrafish (Danio rerio) to determine the function of ET. We cloned and assayed three related proteins from zebrafish, only one of which catalyzed the uptake of ET. RT-PCR analysis revealed that the protein is strongly expressed in the skin, brain, kidney, intestine, and eye. In ETT-knockout animals generated by retroviral insertion into exon 1, ET content was reduced by more than 1000-fold compared to the wild type. Thus, ETT is the sole transporter responsible for uptake of ET into zebrafish. ETT-knockout fish did not exhibit obvious differences in morphology or behavior. In whole-fish homogenates, an increase in 4-hydroxy-2,3-trans-nonenal and malondialdehyde was observed, but only after stress caused by incubation with Pb2+ or Cu2+. Comparison of unstressed fish at the level of small molecules by LC-MS difference shading revealed a 3.8-fold increase in 8-oxoguanine (8-oxo-7,8-dihydroguanine) in the skin of ETT-knockout animals. Our knockout represents a new model for examining the consequences of complete absence of ET. Based on the phenotype observed here, we hypothesize that the specific purpose of ET could be to eliminate singlet oxygen. (C) 2015 Elsevier Inc. All rights reserved.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Pfeiffer, Carolin UNSPECIFIED UNSPECIFIED UNSPECIFIED Bach, Markus UNSPECIFIED UNSPECIFIED UNSPECIFIED Bauer, Tim UNSPECIFIED UNSPECIFIED UNSPECIFIED da Ponte, Julia Campos UNSPECIFIED UNSPECIFIED UNSPECIFIED Schoemig, Edgar UNSPECIFIED UNSPECIFIED UNSPECIFIED Gruendemann, Dirk UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-402754
DOI:	10.1016/j.freeradbiomed.2015.02.026
Journal or Publication Title:	Free Radic. Biol. Med.
Volume:	83
Page Range:	S. 178 - 186
Date:	2015
Publisher:	ELSEVIER SCIENCE INC
Place of Publication:	NEW YORK
ISSN:	1873-4596
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SINGLET MOLECULAR-OXYGEN; INSERTIONAL MUTAGENESIS; GENE-EXPRESSION; DAMAGE; GLUTAMATE; PROFILE; EFFLUX; THIOLS; KIDNEY; CELLS Multiple languages Biochemistry & Molecular Biology; Endocrinology & Metabolism Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/40275