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ToxSci Advance Access originally published online on February 16, 2008
Toxicological Sciences 2008 103(1):68-76; doi:10.1093/toxsci/kfn034
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© The Author 2008. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Deletion of Yeast CWP Genes Enhances Cell Permeability to Genotoxic Agents

Min Zhang*, Yuping Liang{dagger}, Xiaohua Zhang*, Ying Xu*, Heping Dai*,1 and Wei Xiao*,{dagger}

* Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, 430072 China {dagger} Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon SK S7N 5E5, Canada

1 To whom correspondence should be addressed at Institute of Hydrobiology, Chinese Academy of Sciences, 7 Donghu Road South, Wuhan, Hubei, 430072 China. Fax: +86-27-68780123. E-mail: hpdai{at}ihb.ac.cn.

Received December 26, 2007; accepted February 7, 2008


  Abstract

We have previously reported the development of a novel genotoxic testing system based on the transcriptional response of the yeast RNR3-lacZ reporter gene to DNA damage. This system appears to be more sensitive than other similar tests in microorganisms, and is comparable with the Ames test. In an effort to further enhance detection sensitivity, we examined the effects of altering major cell wall components on cell permeability and subsequent RNR3-lacZ sensitivity to genotoxic agents. Although inactivation of single CWP genes encoding cell wall mannoproteins had little effect, the simultaneous inactivation of both CWP1 and CWP2 had profound effects on the cell wall structure and permeability. Consequently, the RNR3-lacZ detection sensitivity is markedly enhanced, especially to high molecular weight compounds such as 4-nitroquinoline-N-oxide (> sevenfold) and phleomycin (> 13-fold). In contrast, deletion of genes encoding representative membrane components or membrane transporters had minor effects on cell permeability. We conclude that the yeast cell wall mannoproteins constitute the major barrier to environmental genotoxic agents and that their removal will significantly enhance the sensitivity of RNR-lacZ as well as other yeast-based genotoxic tests.

Key Words: RNR3-lacZ; yeast; cell wall; mannoprotein; genotoxicity test; sensitivity; permeability.


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