A Novel WRKY-like Protein Involved in Transcriptional Activation of Cyst Wall Protein Genes in Giardia lamblia*
- From the Department of Parasitology, College of Medicine, National Taiwan University, Taipei 100, Taiwan
- 1To whom correspondence should be addressed: Dept. of Parasitology, College of Medicine, National Taiwan University, Taipei, 100 Taiwan. Tel.: 886-2-23123456-88262; Fax: 886-2-23915294; E-mail: chinhsun{at}ntu.edu.tw.
Abstract
Synthesis of a protective cyst wall is required for survival outside of the host and for infection of Giardia lamblia. Little is known of gene regulation of the cyst wall proteins (CWPs) during differentiation into dormant cysts. WRKY homologues constitute a large family of DNA-binding proteins in plants that are involved in several key cellular functions, including disease resistance, stress response, dormancy, and development. A putative wrky gene has been identified in the G. lamblia genome. We found that wrky expression levels increased significantly during encystation. The epitope-tagged WRKY was translocated into the nuclei during encystation. Recombinant WRKY specifically bound to its own promoter and the encystation-induced cwp1 and cwp2 promoters. WRKY contains several key residues for DNA binding, and mutation analysis revealed that its binding sequences are similar to those of the known plant WRKY proteins and that two of them are positive cis-acting elements of the wrky and cwp2 promoters. Overexpression of WRKY increased the cwp1-2 and myb2 mRNA levels, and these gene promoters were bound by WRKY in vivo. Interestingly, the wrky and cwp1-2 genes were up-regulated by ERK1 (extracellular signal-related kinase 1) overexpression, suggesting that WRKY may be a downstream component of the ERK1 pathway. In addition, a WRKY mutant that cannot enter nuclei and an ERK1 mutant lacking the predicted kinase domain showed decreased cwp1-2 gene expression. Our results suggest that the WRKY family has been conserved during evolution and that WRKY is an important transactivator of the cwp1-2 genes during G. lamblia differentiation into dormant cysts.
Footnotes
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↵* This work was supported by National Science Council Grants NSC 94-2320-B-002-093 and NSC 96-2320-B-002-040-MY3 and National Health Research Institutes Grants NHRI-EX96-9510NC, NHRI-EX97-9510NC, and NHRI-EX98-9510NC in Taiwan and was also supported in part by the Department of Medical Research of National Taiwan University Hospital.
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The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. S1.
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↵3 Y. C. Huang and C. H. Sun, unpublished data.
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↵2 The abbreviations used are:
- MAPK
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mitogen-activated protein kinase
- ChIP
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chromatin immunoprecipitation
- RT
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reverse transcription
- HA
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hemagglutinin
- CWP
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cell wall protein.
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- Received April 23, 2009.
