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A high molecular mass zinc transporter MTP 12 forms a functional heteromeric complex with MTP 5 in the Golgi in Arabidopsis thaliana
Author(s) -
Fujiwara Takashi,
Kawachi Miki,
Sato Yori,
Mori Haruki,
Kutsutsumaro,
Hasezawa Seiichiro,
Maeshima Masayoshi
Publication year - 2015
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.13252
Subject(s) - arabidopsis thaliana , bimolecular fluorescence complementation , golgi apparatus , biochemistry , biology , major facilitator superfamily , mutant , saccharomyces cerevisiae , microbiology and biotechnology , heterologous expression , transport protein , chemistry , yeast , endoplasmic reticulum , gene , recombinant dna
Zinc (Zn) is an essential micronutrient required for plant growth and development. In Arabidopsis thaliana , several families of Zn transporters engaged in Zn import, export and intracellular compartmentalization play important roles in Zn homeostasis. We describe a novel Zn transporter, A. thaliana metal tolerance protein 12 (At MTP 12), which belongs to the cation diffusion facilitator family. At MTP 12 is predicted to consist of 798 amino acids and have 14 transmembrane segments. The expression of At MTP 12 in suspension‐cultured cells was not affected by Zn deficiency or excess. Heterologous expression in a mutant of budding yeast ( Saccharomyces cerevisiae ) that lacks Msc2p, an orthologue of At MTP 12, revealed that At MTP 12 complements the growth phenotype of the msc2 mutant when At MTP 5t1, one of the splicing variants of At MTP 5, is coexpressed. Transient expression of At MTP 12‐fused green fluorescent protein in A. thaliana mesophyll protoplasts demonstrated that At MTP 12 is localized to the Golgi apparatus. Moreover, At MTP 12 and At MTP 5t1 interact in the Golgi, as determined by a bimolecular fluorescence complementation assay. These results suggest that At MTP 12 forms a functional complex with At MTP 5t1 to transport Zn into the Golgi. Database Nucleotide sequence data for full‐length of AtMTP12 is available in the DDBJ/EMBL/GenBank database under accession number AB986563 .