Abstract
Wor1 (white-opaque switching regulator 1) is a master regulator of the white-opaque switching in Candida albicans, an opportunistic human fungal pathogen, and is associated with its pathogenicity and commensality. Wor1 contains a conserved DNA-binding region at the N-terminus, consisting of two conserved segments (WOPRa and WOPRb) connected by a non-conserved linker that can bind to specific DNA sequences of the promoter regions and then regulates the transcription. Here, we report the crystal structure of the C. albicans Wor1 WOPR segments in complex with a double-stranded DNA corresponding to one promoter region of WOR1. The sequentially separated WOPRa and WOPRb are structurally interwound together to form a compact globular domain that we term the WOPR domain. The WOPR domain represents a new conserved fungal-specific DNA-binding domain which uses primarily a conserved loop to recognize and interact specifically with a conserved 6-bp motif of the DNA in both minor and major grooves. The protein-DNA interactions are essential for WOR1 transcriptional regulation and white-to-opaque switching. The structural and biological data together reveal the molecular basis for the recognition and binding specificity of the WOPR domain with its specific DNA sequences and the function of Wor1 in the activation of transcription.
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Acknowledgements
We thank the staff members at BL17U of Shanghai Synchrotron Radiation Facility (SSRF), China, for technical support in diffraction data collection and other members of our groups for helpful discussion. This work was supported by the National Natural Science Foundation of China (31370105 and 31230017), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB08010300), and the Ministry of Science and Technology of China (2011CB966301 and 2010CB912103).
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Supplementary information
Supplementary information, Figure S1
Sequence comparison of the WOPRb domain from different fungal species. (PDF 453 kb)
Supplementary information, Figure S2
Binding of the Wor1 WOPR domain with dsDNA of different sequences. (PDF 85 kb)
Supplementary information, Figure S3
Crystal structure of the WOPR-17bp dsDNA complex. (PDF 103 kb)
Supplementary information, Figure S4
Comparison of the WOPR-13bp dsDNA complex and the WOPR-17bp dsDNA complex. (PDF 116 kb)
Supplementary information, Figure S5
Comparison of the CaWor1 WOPR domain with the NAC domain. (PDF 144 kb)
Supplementary information, Figure S6
Sequence comparison of the WOPR domain between CaWor1 and CaPth2. (PDF 40 kb)
Supplementary information, Table S1
Interactions between theWOPR domain and the dsDNA (PDF 26 kb)
Supplementary information, Table S2
Binding affinity of WOPR mutations with dsDNA (PDF 9 kb)
Supplementary information, Table S3
dsDNA used in the EMSA (PDF 13 kb)
Supplementary information, Data S1
Materials and Methods (PDF 19 kb)
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Zhang, S., Zhang, T., Yan, M. et al. Crystal structure of the WOPR-DNA complex and implications for Wor1 function in white-opaque switching of Candida albicans. Cell Res 24, 1108–1120 (2014). https://doi.org/10.1038/cr.2014.102
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DOI: https://doi.org/10.1038/cr.2014.102