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Engineering expression of the heavy metal transporter MerC in Saccharomyces cerevisiae for increased cadmium accumulation
Authors:Masako Kiyono  Kiyomi Miyahara  Yuka Sone  Hidemitsu Pan-Hou  Shimpei Uraguchi  Ryosuke Nakamura  Kou Sakabe
Affiliation:(1) Department of Public Health and Molecular Toxicology, School of Pharmacy, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan;(2) Faculty of Pharmaceutical Sciences, Setsunan University, 45-1 Nagaotoge-cho, Hirakata, Osaka 573-0101, Japan;(3) Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
Abstract:The merC gene from the Tn21-encoded mer operon has potential uses as a molecular tool for bioremediation. It was overexpressed as the fusion proteins MerC-Sso1p or MerC-Vam3p in Saccharomyces cerevisiae. Green fluorescent protein (GFP)-MerC-Sso1p fusion proteins located primarily in the plasma membrane, although some protein was detected in the endoplasmic reticulum. In contrast, GFP-MerC-Vam3p was expressed in the vacuolar membranes. These results suggest that yeast Sso1p and Vam3p are essential for targeting molecules to the plasma and vacuolar membranes, respectively. Significantly more cadmium ions were accumulated by yeast cells expressing MerC-Sso1p than with MerC-Vam3p or control cells. These results suggest that expression of MerC in the plasma membrane may be a particularly promising strategy for improving accumulation of cadmium in yeast.
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