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Zebrafish Ubc13 is required for Lys63-linked polyubiquitination and DNA damage tolerance

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Abstract

Ubiquitination is an important post-translational protein modification that functions in diverse cellular processes of all eukaryotic organisms. Conventional Lys48-linked poly-ubiquitination leads to the degradation of specific proteins through 26S proteasomes, while Lys63-linked polyubiquitination appears to regulate protein activities in a non-proteolytic manner. To date, Ubc13 is the only known ubiquitin-conjugating enzyme capable of poly-ubiquitinating target proteins via Lys63-linked chains, and this activity absolutely requires a Ubc variant (Uev or Mms2) as a co-factor. However, Lys63-linked poly-ubiquitination and error-free DNA damage tolerance in zebrafish are yet to be defined. Here, we report molecular cloning and functional characterization of two zebrafish ubc13 genes, ubc13a and ubc13b. Analysis of their genomic structure, nucleotide and protein sequence indicates that the two genes are highly conserved during evolution and derived from whole genome duplication. Zebrafish Ubc13 proteins are able to physically interact with yeast or human Mms2 and both zebrafish ubc13 genes are able to functionally complement the yeast ubc13 null mutant for spontaneous mutagenesis and sensitivity to DNA damaging agents. In addition, upon DNA damage, the expression of zebrafish ubc13a and ubc13b is induced during embryogenesis and zebrafish Ubc13 is associated with nuclear chromatin. These results suggest the involvement of Lys63-linked poly-ubiquitylation in DNA damage response in zebrafish.

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Acknowledgments

We thank various laboratories for kindly providing reagents and plasmids, Drs. Z. Zhu and H. Dai for their valuable comments and suggestions, and all other members in Cui and Xiao Laboratories for helpful suggestions and technical assistance. This work was supported by grants from the National Basic Research Program of China (#2009CB941200) and the Canadian Institutes of Health Research MOP-53240 to WX.

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Authors and Affiliations

  1. Institute of Hydrobiology, Chinese Academy of Sciences, 7 Donghu Rd., Wuhan, 430072, Hubei, People’s Republic of China

    Jie Li, Qing Li & Zongbin Cui

  2. Department of Microbiology and Immunology, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, S7N 5E5, Canada

    Rui Wen, Parkeer Andersen, Yuping Liang & Wei Xiao

  3. Graduate University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Jie Li

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  1. Jie Li
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  2. Rui Wen
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  4. Yuping Liang
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  5. Qing Li
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  6. Wei Xiao
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Correspondence to Wei Xiao or Zongbin Cui.

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Li, J., Wen, R., Andersen, P. et al. Zebrafish Ubc13 is required for Lys63-linked polyubiquitination and DNA damage tolerance. Mol Cell Biochem 343, 173–182 (2010). https://doi.org/10.1007/s11010-010-0511-9

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  • DOI: https://doi.org/10.1007/s11010-010-0511-9

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