Vivo-Morpholinos: Same oligos, from cultures to critters -


Combined zebrafish-yeast chemical-genetic screens reveal gene–copper-nutrition interactions that modulate melanocyte pigmentation
Hironori Ishizaki, Michaela Spitzer, Jan Wildenhain, Corina Anastasaki, Zhiqiang Zeng, Sonam Dolma, Michael Shaw, Erik Madsen, Jonathan Gitlin, Richard Marais, Mike Tyers, E. Elizabeth Patton


Hypopigmentation is a feature of copper deficiency in humans, as caused by mutation of the copper (Cu2+) transporter ATP7A in Menkes disease, or an inability to absorb copper after gastric surgery. However, many causes of copper deficiency are unknown, and genetic polymorphisms might underlie sensitivity to suboptimal environmental copper conditions. Here, we combined phenotypic screens in zebrafish for compounds that affect copper metabolism with yeast chemical-genetic profiles to identify pathways that are sensitive to copper depletion. Yeast chemical-genetic interactions revealed that defects in intracellular trafficking pathways cause sensitivity to low-copper conditions; partial knockdown of the analogous Ap3s1 and Ap1s1 trafficking components in zebrafish sensitized developing melanocytes to hypopigmentation in low-copper environmental conditions. Because trafficking pathways are essential for copper loading into cuproproteins, our results suggest that hypomorphic alleles of trafficking components might underlie sensitivity to reduced-copper nutrient conditions. In addition, we used zebrafish-yeast screening to identify a novel target pathway in copper metabolism for the small-molecule MEK kinase inhibitor U0126. The zebrafish-yeast screening method combines the power of zebrafish as a disease model with facile genome-scale identification of chemical-genetic interactions in yeast to enable the discovery and dissection of complex multigenic interactions in disease-gene networks.


  • * These authors contributed equally to this work

  • Present address: The Hospital for Sick Children, Developmental Stem Cell Biology, TMDT Building, 13-601G, 101 College Street, Toronto, Ontario, M5G 1L7, Canada


    The authors declare no financial, personal or professional associations that interfere with the objectivity of this study.


    H.I. and M.S. conceived, designed and performed experiments, and J.W., C.A., Z.Z., M.S., E.M. and E.E.P. performed experiments and analyzed data. S.D., J.G. and R.M. contributed to experimental design and provided reagents. M.T. and E.E.P. conceived and designed experiments, and wrote the paper.


    Supplementary material for this article is available at

  • Received April 2, 2010.
  • Accepted May 27, 2010.
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