Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jan 26;24(3):2412.
doi: 10.3390/ijms24032412.

Functional Characterization of Potato UBC13- UEV1s Genes Required for Ubiquitin Lys63 Chain to Polyubiquitination

Weigang Liu  1   2 , Xun Tang  1   3 , Xue Fu  1   2 , Huanhuan Zhang  1   2 , Cunlan Zhu  1   3 , Ning Zhang  1   3 , Huaijun Si  1   3
Affiliations

Functional Characterization of Potato UBC13- UEV1s Genes Required for Ubiquitin Lys63 Chain to Polyubiquitination

Weigang Liu et al. Int J Mol Sci. .

Abstract

Ubiquitin-conjugating enzymes (E2s/UBC) are components of the ubiquitin proteasome system (UPS), and the ubiquitin-conjugating enzyme variant (UEV) is one of E2s (ubiquitin-conjugating enzymes, UBC) subfamily. The UEVs and UBC13 play an auxiliary role in mediating Lys63-linked polyUb chain assembly, which is correlated with target protein non-proteolytic functions, such as DNA repair or response to stress. However, the collaborative mechanism of StUBC13 (homologue of AtUBC13) and StUEVs (the UEVs in potato) involved in potato are not fully understood understood. Here, we identified two StUBC13 and seven StUEVs from potato genome. We analyzed protein motif and conserved domain, gene structure, phylogenetic features, cis-acting elements of StUBC13 and StUEVs. Subsequently, we screened StUBC13 partners protein and verified interaction between StUBC13 and StUEVs using yeast two-hybrid, split luciferase complementation (SLC) and bimolecular fluorescence complementation (BiFC) approach. The expression profile and qRT-PCR analysis suggested that StUBC13 and StUEVs gene exhibited a tissue-specific expression and were induced by different stress. Overall, this investigative study provides a comprehensive reference and view for further functional research on StUBC13 and StUEV1s in potato.

Keywords: UBC13; UEVs; gene expression; potato; protein interaction.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Characterization analysis of UEVs gene family. (a) Phylogenetic relationships and classification of UEVs gene family. (b) The three-dimensional structure of UEVs conserved domain. (c) Protein sequence alignment of VPS23 subgroups in Arabidopsis and potato. (d) Protein sequence alignment of UEV1s subgroups from Oryza sativa, Arabidopsis thaliana, Brachypodium distachyon, and Solanum tuberosum.
Figure 2
Figure 2
The cis-acting element, conserved motif, and gene structure analysis of UEVs. (a) The cis-acting element analysis of StUEVs genes in the promoter region. (b) The motif composition analysis of StUEVs. (c) The gene structure analysis of StUEVs.
Figure 3
Figure 3
The characterization of StUBC13. (a) Phylogenetic relationships of UBC13 from Oryza sativa, Arabidopsis thaliana, Brachypodium distachyon, Solanum lycopersicum, and Solanum tuberosum. (b) Gene structure analysis of UBC13. (c) Protein sequence alignment of UBC13. * indicates that this residue is conserved.
Figure 4
Figure 4
StUBC13 interacts with multiple proteins. (a) The verification of StUBC13 and multiple protein interactions by one-to-one Y2H. (b) The interaction between StRGLG1, StUEV1B and StUBC13 in BiFC assay. (c) The interaction between StRGLG1, StUEV1B and StUBC13 in SLC assay.
Figure 5
Figure 5
Subcellular localization of StUBC13 and its partners. The red arrows indicate the nucleus, the blue arrows the cell membrane, and the yellow arrows indicate membrane.
Figure 6
Figure 6
The expression patterns of StUEVs and StUBC13. (a) The tissue-specific heatmap of StUEVs and StUBC13 in DM. (b) The tissue-specific heatmap of StUEVs and StUBC13 in RH. (c) The expression patterns of StUEVs and StUBC13 under different treatments in DM. (d) The expression patterns of StUEVs and StUBC13 in “Atlantic” and “Qingshu” during the flowering stage under drought stress.
Figure 7
Figure 7
The qRT-PCR analysis of StUEVs and StUBC13 in different potato tissues. Different small letters indicate statistical differences of tissues when assessed using Duncan’s multiple range test (p < 0.05, n = 3).
Figure 8
Figure 8
The qRT-PCR analysis of StUEVs and StUBC13 under different stress. Different small letters indicate the statistical differences of tissues when assessed using Duncan’s multiple range test (p < 0.05, n = 3).
See this image and copyright information in PMC

References

    1. Xu F.Q., Xue H.W. The ubiquitin-proteasome system in plant responses to environments. Plant Cell Environ. 2019;42:2931–2944. doi: 10.1111/pce.13633. - DOI - PubMed
    1. Komander D., Rape M. The Ubiquitin Code. Annu. Rev. Biochem. 2012;81:203–229. doi: 10.1146/annurev-biochem-060310-170328. - DOI - PubMed
    1. Wickliffe K.E., Williamson A., Meyer H.-J., Kelly A., Rape M. K11-linked ubiquitin chains as novel regulators of cell division. Trends Cell Biol. 2011;21:656–663. doi: 10.1016/j.tcb.2011.08.008. - DOI - PMC - PubMed
    1. Cheng M.-C., Kuo W.-C., Wang Y.-M., Chen H.-Y., Lin T.-P. UBC18 mediates ERF1 degradation under light–dark cycles. New Phytol. 2017;213:1156–1167. doi: 10.1111/nph.14272. - DOI - PubMed
    1. Saeki Y., Kudo T., Sone T., Kikuchi Y., Yokosawa H., Toh-e A., Tanaka K. Lysine 63-linked polyubiquitin chain may serve as a targeting signal for the 26S proteasome. EMBO J. 2009;28:359–371. doi: 10.1038/emboj.2008.305. - DOI - PMC - PubMed

MeSH terms

LinkOut - more resources