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Accumulation of Ag and Cu in Amanita strobiliformis and characterization of its Cu and Ag uptake transporter genes AsCTR2 and AsCTR3
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SYSNO ASEP 0458673 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Accumulation of Ag and Cu in Amanita strobiliformis and characterization of its Cu and Ag uptake transporter genes AsCTR2 and AsCTR3 Author(s) Beneš, V. (CZ)
Hložková, K. (CZ)
Matěnová, M. (CZ)
Borovička, Jan (UJF-V) RID, ORCID, SAI
Kotrba, P. (CZ)Number of authors 5 Source Title Biometals. - : Springer - ISSN 0966-0844
Roč. 29, č. 2 (2016), s. 249-264Number of pages 16 s. Publication form Print - P Language eng - English Country NL - Netherlands Keywords heavy metals ; metal uptake ; Metallothionein ; copper transporter protein family ; Ectomycorrhizal fungi Subject RIV CB - Analytical Chemistry, Separation R&D Projects GAP504/11/0484 GA ČR - Czech Science Foundation (CSF) LM2015056 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UJF-V - RVO:61389005 UT WOS 000372165700007 EID SCOPUS 84957611852 DOI 10.1007/s10534-016-9912-x Annotation Macrofungi can accumulate in their sporocarps remarkably high concentrations of Cu and Ag. We have previously demonstrated that the non-essential Ag is in the ectomycorrhizal, Ag-hyperaccumulating Amanita strobiliformis sequestered by 3.4-kDa metallothioneins (MTs) produced as AsMT1a, 1b and 1c isoforms. Here, we describe two populations of wild-grown A. strobiliformis sporocarps, which showed certain correlation between the concentrations of accumulated Ag (284 +/- A 64 and 67 +/- A 15 mg kg(-1)) and Cu (76 +/- A 13 and 30 +/- A 12 mg kg(-1)), suggesting that an overlap may exist in the cell biology of Ag and Cu in this species. Metal speciation analysis revealed that the intracellular Cu in the sporocarps of both populations was, like Ag, associated with the 3.4-kDa MTs. A search of A. strobiliformis transcriptome for sequences encoding proteins of the Cu transporter (CTR) family identified four AsCTR cDNAs, which were, like AsMT1s, confirmed in both populations. The predicted AsCTR proteins showed homology to vacuolar (AsCTR1 and AsCTR4) and plasma membrane (AsCTR2 and AsCTR3) CTRs. Heterologous expression of AsCTR2, AsCTR3 and their translational fusions with green fluorescent protein (GFP) in Cu uptake-deficient S. cerevisiae indicated that both AsCTRs are functional Cu and Ag uptake transporters: recombinant genes complemented growth defects and increased Cu and Ag uptake rates in yeasts and the GFP-tagged protein localized to the cell periphery. Site directed mutagenesis revealed the importance of the conserved-among-CTRs M-X-3-M motif for the AsCTR2- and AsCTR3-mediated transport of both Cu and Ag. These results provide the first evidence that fungal CTRs can recognize Ag for transport. Workplace Nuclear Physics Institute Contact Markéta Sommerová, sommerova@ujf.cas.cz, Tel.: 266 173 228 Year of Publishing 2017
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