title: Copper induces Cu-ATPase ATP7A mRNA in a fish cell line, SAF1.
abstract: Copper transporting ATPase, ATP7A, is an ATP dependent copper pump present in all vertebrates, critical for the maintenance of intracellular and whole body copper homeostasis. Effects of copper treatment on ATP7A gene expression in fibroblast cells (SAF1) of the sea bream (Sparus aurata) were investigated by qRT-PCR and by a medium density microarray from a closely related species, striped sea bream (Lithognathus mormyrus). To discriminate between the effects of Cu and other metals, SAF1 cells were exposed to sub-toxic levels of Cu, Zn and Cd. Expression of Cu homeostasis genes copper transporter 1 (CTR1), Cu ATPase (ATP7A), Cu chaperone (ATOX1) and metallothionein (MT) together with the oxidative stress markers glutathione reductase (GR) and Cu/Zn superoxide dismutase (CuZn/SOD) were measured 0, 4 and 24 hours post-exposure by qRT-PCR. Microarray was conducted on samples from 4 hours post Cu exposure. Cu, Zn and Cd increased MT and GR mRNA levels, while only Cu increased ATP7A mRNA levels. Microarray results confirmed the effects of Cu on ATP7A and MT and in addition showed changes in the expression of genes involved in protein transport and secretion. Results suggest that ATP7A may be regulated at the transcriptional level directly by Cu and by a mechanism that is different from that exerteted by metals on MT genes.
authors.affiliation: Institute of Aquaculture, University of Stirling, UK. matteo.minghetti@stir.ac.uk
authors.forename: Matteo
authors.initials: M
authors.lastname: Minghetti
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Leaver
authors.forename: John B
authors.initials: JB
authors.lastname: Taggart
authors.forename: Elisa
authors.initials: E
authors.lastname: Casadei
authors.forename: Meirav
authors.initials: M
authors.lastname: Auslander
authors.forename: Moshe
authors.initials: M
authors.lastname: Tom
authors.forename: Stephen G
authors.initials: SG
authors.lastname: George
mesh_terms: D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D027682:Cation Transport Proteins / Q000096:biosynthesis / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019327:Copper Sulfate / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D046228:Microarray Analysis; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D021541:Sea Bream / Q000502:physiology*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D027682:Cation Transport Proteins; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical; D010088:Oxidoreductases; D000251:Adenosine Triphosphatases; D019327:Copper Sulfate