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. 2020 Mar 21;12(3):346. doi: 10.3390/v12030346

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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TRIM21 mediates the K48-linked ubiquitination of HBV DNA Pol at K260 and K283. (A) A schematic diagram of the lysine point mutations of ubiquitin. (B) Overexpression of HBV DNA Pol and TRIM21 in Huh7 cells cotransfected with HA-ubiquitin, Ub-K48R or Ub-K63R mutants. A ubiquitination assay and Western blot were used to detect the ubiquitination and protein level of HBV DNA Pol. (C) Schematic of the potential ubiquitination sites of HBV DNA Pol. (D) Huh7 cells were transfected with FLAG-HBV DNA Pol or its lysine point mutants, and ubiquitination assays were used to analyze the ubiquitination level of HBV DNA Pol and its lysine mutants. (E) TRIM21 and FLAG-HBV DNA Pol or its lysine mutants were cotransfected into Huh7 cells, and the effect of TRIM21 on the ubiquitination level of HBV DNA Pol and its lysine mutants was detected by ubiquitination assay. Data are representative of three independent experiments with three replicates each.