Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation

نویسندگان

  • Lavinia Liliana Ruta
  • Ya-Fen Lin
  • Ralph Kissen
  • Ioana Nicolau
  • Aurora Daniela Neagoe
  • Simona Ghenea
  • Atle M Bones
  • Ileana Cornelia Farcasanu
چکیده

In this study we engineered yeast cells armed for heavy metal accumulation by targeting plant metallothioneins to the inner face of the yeast plasma membrane. Metallothioneins (MTs) are cysteine-rich proteins involved in the buffering of excess metal ions, especially Cu(I), Zn(II) or Cd(II). The cDNAs of seven Arabidopsis thaliana MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four Noccaea caerulescens MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) were each translationally fused to the C-terminus of a myristoylation green fluorescent protein variant (myrGFP) and expressed in Saccharomyces cerevisiae cells. The myrGFP cassette introduced a yeast myristoylation sequence which allowed directional targeting to the cytosolic face of the plasma membrane along with direct monitoring of the intracellular localization of the recombinant protein by fluorescence microscopy. The yeast strains expressing plant MTs were investigated against an array of heavy metals in order to identify strains which exhibit the (hyper)accumulation phenotype without developing toxicity symptoms. Among the transgenic strains which could accumulate Cu(II), Zn(II) or Cd(II), but also non-canonical metal ions, such as Co(II), Mn(II) or Ni(II), myrGFP-NcMT3 qualified as the best candidate for bioremediation applications, thanks to the robust growth accompanied by significant accumulative capacity.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Characterization of Phosphate Membrane Transport in Saccharomyces cerevisiae CEN.PK113-5D under Low-Phosphate Conditions Using Aerobic Continuous Culture

Two different growth media, namely complex and defined media, were used to examine establishment of steady-state conditions in phosphate-limited culture system of Saccharomyces cerevisiae CEN.PK113-5D strain. Using the defined growth medium, it was possible to obtain steady state condition in the continuous culture. The effect of phosphate concentration on the growth of S. cerevisiae in pho...

متن کامل

Heavy metal regulation of plasma membrane H+-ATPase gene expression in halophyte Aeluropus littoralis

The present study was conducted to find the effect of three heavy metals, Ag, Hg and Pb on the expression level of a gene encoding plasma membrane H+-ATPase in Aeluropus littoralis. The experiment was laid out in a completely random design with three replications. The expression of the main gene was normalized to the expression of the housekeeping gene actin. Two 259 and 187 bp fragments were a...

متن کامل

The Mechanism of Chromium Biosorption by Saccharomyces Cerevisiae

The Biosorption property of S. cerevisiae for chromium uptake was investigated in an immobilized cell bioreactor. Saw dust was utilized as the solid bed in the reactor. Adsorption of S. cerevisiae on saw dust obeys a first order reaction kinetic up to 6 hours. The immobilized biomass particles are porous and exist in the new generation of biological adsorbent. Chromium biosorption was studied i...

متن کامل

TcOPT3, a Member of Oligopeptide Transporters from the Hyperaccumulator Thlaspi caerulescens, Is a Novel Fe/Zn/Cd/Cu Transporter

BACKGROUND Thlaspi caerulescens is a natural selected heavy metal hyperaccumulator that can not only tolerate but also accumulate extremely high levels of heavy metals in the shoots. Thus, to identify the transportors involved in metal long-distance transportation is very important for understanding the mechanism of heavy metal accumulation in this hyperaccumulator. METHODOLOGY/PRINCIPAL FIND...

متن کامل

Toxic Effects of Heavy Metals on Plant Growth and Metal Accumulation in Maize (Zea maysL.)

Background: A pot experiment was conducted to determine the toxic effects of some heavy metals on the plant growth and seed yield of maize (Zea mays L.). Materials and Metheds: Heavy metals Mn, Pb, Cd, Cr and Co individually and in combinations were added as chloride salts in solutions to the pots before sowing. The test plants for were harvested after 80 days of germination and evaluated f...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 12  شماره 

صفحات  -

تاریخ انتشار 2017