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Issue 6, 2011
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A novel ubiquitin binding mode in the S. cerevisiaetranslesion synthesis DNA polymerase η

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Abstract

The ubiquitin binding zinc finger (UBZ) domain in the C-terminal portion of Polη has been found to interact with ubiquitin. However, the affinity between the Polη UBZ and ubiquitin was shown to be low with a previously reported Kd of 73–81 μM. This low-affinity binding between Polη UBZ and ubiquitin has been difficult to reconcile with its presumed role in translesion synthesis as suggested by genetic and cell biology studies. In this work, we constructed a minimal S. cerevisiae Polη UBZ domain and probed the Polη UBZ-ubiquitin interaction using a surface plasmon resonance (SPR) technique. Our quantitative binding data between the wild-type or mutant Polη UBZ and ubiquitin revealed an interesting divergence between the Polη UBZ from S. cerevisiae and humans. Moreover, we found that the C-terminal portion of yeast Polη (amino acid 515–632) binds ubiquitin with a much higher affinity than the minimal UBZ domain. Further, distinct ubiquitin-binding kinetics were observed for the C-terminal portion of Polη and the isolated UBZ domain. This observation raised the interesting possibility that the Polη C-terminal portion binds ubiquitin in a novel mode that affords higher affinity. Our findings have broader implication in understanding the generally weak interaction between the known ubiquitin-binding domains and ubiquitin.

Graphical abstract: A novel ubiquitin binding mode in the S. cerevisiaetranslesion synthesis DNA polymerase η

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Supplementary files

Article information


Submitted
31 Dec 2010
Accepted
15 Mar 2011
First published
11 Apr 2011

Mol. BioSyst., 2011,7, 1874-1882
Article type
Paper

A novel ubiquitin binding mode in the S. cerevisiaetranslesion synthesis DNA polymerase η

Y. Ai, J. Wang, R. E. Johnson, L. Haracska, L. Prakash and Z. Zhuang, Mol. BioSyst., 2011, 7, 1874 DOI: 10.1039/C0MB00355G

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