Cadmium hyperaccumulating mushroom Cystoderma carcharias has two metallothionein isoforms usable for cadmium and copper storage.

ABSTRACT

Cystoderma carcharias is one of the few macrofungal species that can hyperaccumulate Cd. As we have previously documented in C. carcharias collected from a smelter-polluted area, it stores 40% of Cd and nearly 90% of Cu in sporocarps in complex(es) of identical size. In this paper we examined whether metallothionein (MT) peptides that bind Cd and Cu through cysteinyl-thiolate bonds were associated with the metals in these complexes. Screening of a sporocarp cDNA expression library in yeasts allowed the identification of two transcripts, CcMT1 and CcMT2, encoding functional 34-amino acid (AA) MTs sharing 56% identity and appearing to be encoded by duplicate genes. CcMT1 conferred reasonable tolerance to Cu and a substantially higher tolerance to Cd than CcMT2, while CcMT2 clearly protected the yeasts better against Cu toxicity. While size-exclusion chromatography revealed that CcMT1 was contained in all Cd/Cu complexes isolated from wild grown sporocarps, CcMT2 was detected in a much narrower subset of the fractions. The striking difference between the CcMTs is that CcMT1 lacks the third metal-biding cysteinyl (C) within an otherwise highly conserved-in-agaricomycetes-MTs C-AA4-C-AA-C-AA3-C-AA-C-AA4-C-AA-C motif. The elimination of the corresponding cysteinyl in CcMT2 only reduced the Cu-tolerant phenotype in yeasts to the levels observed with CcMT1. Altogether, these results indicate that CcMT2 is rather adjusted to perform Cu-related tasks and point to CcMT1 as the ligand for the storage of both Cd and Cu in C.carcharias, which is the first macrofungal species in which the potential of MT in Cd handling can be seen.