Abstract
Auxilin is a J-domain-containing defense-related protein involved in clathrin-mediated endocytosis (CME) and acts on many organisms including yeast, Caenorhabditis elegans, Drosophila, Arabidopsis thaliana and rice. Auxilin regulates biological processes such as signal transduction, antibacterial defense, and stress resistance. To investigate the function of Auxilin gene in response to plant pathogen infection, the full-length of potato (Solanum tuberosum L.) Auxilin gene named StAL1 was cloned. Bioinformatics analysis of the gene revealed that StAL1 consists of 4828 bp, including a 4380 bp open reading frame that encodes a protein with 1443 amino acids. StAL1 is highly similar to Auxilin-like protein in potato and other Solanaceae species. Based on the qRT-PCR assays tissue expression pattern analysis revealed that StAL1 constitutively expressed in potato. StAL1 was induced by Ralstonia solanacearum species complex (RSSC) and up-regulated to varying extents by specific concentrations of exogenous phytohormones, including salicylic acid (SA), methyl jasmonate (MeJA) and abscisic acid (ABA). Examination of tissue localization suggested that StAL1 was primarily expressed in specific tissues (the phloem of the stem and leaf vascular systems). Furthermore, transient expression analysis showed the StAL1-GFP fusion protein was localized in the nucleus of Nicotiana benthamiana leaves cells. More importantly, auxilin is involved in plant immune response and disease resistance as an immune receptor binding protein. The results of the study should lay down the foundation for future research on the functional characterization of the Auxilin in potato plants.
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REFERENCES
Ahle, S. and Ungewickell, E., auxilin, a newly identified clathrin-associated protein in coated vesicles from bovine brain, J. Cell. Biol., 1990, vol. 111, p. 19.
Lemmon, S.K., Clathrin uncoating: auxilin comes to life, Curr. Biol., 2001, vol. 11, p. 49.
Schröder, S., Morris, S.A., Knorr, R., Plessmann, U., Weber, K., Nquyen, G.V., and Ungewickell, E., Primary structure of the neuronal clathrin-associated protein auxilin and its expression in bacteria, Eur. J. Biochem., 1995,vol. 228, p. 297.
Kalli, A.C., Morgan, G., and Sansom, M.S., Interactions of the auxilin-1PTEN-like domain with model membranes result in nanoclustering of phosphatidylinositol phosphates, Biophys. J., 2013, vol. 105, p. 137.
Jiang, J., Maes, E.G., Taylor, A.B., Wang, L., Hinck, A.P., Lafer, E.M., and Sousa, R., Structural basis of J cochaperone binding and regulation of Hsp70, Mol. Cell, 2007, vol. 28, p. 422.
Eisenberg, E. and Greene, L.E., Multiple roles of auxilin and hsc70 in clathrin-mediated endocytosis, Traffic, 2007, vol. 8, p. 640.
Suetsugu, N., Kagawa, T., and Wada, M., An auxilin-like J-domain protein, JAC1, regulates phototropin-mediated chloroplast movement in Arabidopsis, Plant Physiol., 2005, vol. 139, p. 151.
Park, C.J., Wei, T., Sharma, R., and Ronald, P.C., Overexpression of rice auxilin-like protein, XB21, induces necrotic lesions, up-regulates endocytosis-related genes, and confers enhanced resistance to Xanthomonas oryzae pv. oryzae, Rice, 2017, vol. 10, p. 27.
Takano, A., Suetsugu, N., Wada, M., and Kohda, D., Crystallographic and functional analyses of J-domain of JAC1 essential for chloroplast photorelocation movement in Arabidopsis thaliana, Plant Cell Physiol., 2010, vol. 51, p. 1372.
Dauwerse, J.G., Dixon, J., Seland, S., Ruivenkamp, C.A., Van, H.A., Hoefsloot, L.H., Peters, D.J., Boers, A.C., Daumer-Haas, C., Maiwald, R., Zweier, C., Kerr, B., Cobo, A.M., Toral, J.F., Hoogeboom, A.J., et al., Mutations in genes encoding subunits of RNA polymerases I and III cause Treacher Collins syndrome, Nat. Genet., 2011, vol. 43, p. 20.
Ezaki, B., Kiyohara, H., Matsumoto, H., and Nakashima, S., Overexpression of an auxilin-like gene (F9E10.5) can suppress Al uptake in roots of Arabidopsis, J. Exp. Bot., 2007, vol. 58, p. 497.
He, L.Y., Sequeira, L., and Kelman, A., Characteristics of strains of Pseudomonas solanacearum from China, Plant Dis., 1983, vol. 67, p. 1357.
Ni, X.M., Tian, Z.D., Liu, J., Song, B.T., Li, J.C., Shi, X.L., and Xie, C.H., StPUB17, a novel potato UND/PUB/ARM repeat type gene, is associated with late blight resistance and NaCl stress, Plant Sci., 2010, vol. 178, p. 158.
Kong, C.Y., Luo, Y.P., Duan, T.T., Xue, Z., Gao, X.D., Zhao, X., and Gao, G., Potato remorin gene StREMa4 cloning and its spatiotemporal expression pattern under Ralstonia solanacearum and plant hormones treatment, Phytoparasitica, 2016, vol. 44, p. 575.
Gao, G., Ren, C.H., Jin, L.P., Xie, K.Y., and Qu, D.Y., Cloning, expression and characterization of a non-specific lipid transfer protein gene from potato, Acta Agron. Sin., 2008, vol. 34, p. 1510.
Horton, P., Park, K.J., Obayashi, T., Fujita, N., Harada, H., Adams-Collier, C.J., and Nakai, K., WOLF PSORT: protein localization predictor, Nucleic Acids Res., 2007, vol. 35, p. W585.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S., MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods, Mol. Biol. Evol., 2011, vol. 28, p. 2731.
Gao, G., Jin, L.P., Xie, K.Y., and Qu, D.Y., The potato StLTPa7 gene displays a complex Ca2+-associated pattern of expression during the early stage of potato–Ralstonia solanacearum interaction, Mol. Plant Pathol., 2009, vol. 10, p. 15.
Khan, M.I., Syeed, S., Nazar, R., and Anjum, N.A., An insight into the role of salicylic acid and jasmonic acid in salt stress tolerance, in Phytohormones and Abiotic Stress Tolerance in Plants, Berlin: Springer-Verlag, 2012, p. 277.
Adamowski, M., Narasimhan, M., Kania, U., Glanc, M., De, J.G., and Friml, J., A functional study of auxilin-like1 and 2, two putative clathrin uncoating factors in Arabidopsis, Plant Cell, 2018, vol. 30, p. 700.
Du, Y., Tejos, R., Beck, M., Himschoot, E., Li, H., Robatzek, S., Vanneste, S., and Friml, J., Salicylic acid interferes with clathrin-mediated endocytic protein trafficking, Proc. Natl. Acad. Sci. U.S.A., 2013, vol. 110, p. 7946.
Jelenska, J., Yao, N., Vinatzer, B.A., Wright, C.M., Brodsky, J.L., and Greenberg, J.T., A J domain virulence effector of Pseudomonas syringae remodels host chloroplasts and suppresses defenses, Curr. Biol., 2007, vol. 17, p. 499.
Ye, S.F., Yu, S.W., Shu, L.B., Wu, J.H., Wu, A.Z., and Luo, L.J., Expression profile analysis of 9 heat shock protein genes throughout the life cycle and under abiotic stress in rice, Mol. Biol., 2012, vol. 57, p. 336.
Zhang, Z. N., Wang, B., Sun, D.M., and Deng, X., Molecular cloning and differential expression of sHSP gene family; members from the resurrection plant Boea hygrometrica in response to abiotic stresses, Biologia, 2013, vol. 68, p. 651.
Eun, S.H., Banks, S.M., and Fischer, J.A., Auxilin is essential for Delta signaling, Development, 2008, vol. 135, p. 1089.
Zhao, X., Greener, T., Al-Hasani, H., Cushman, S.W., Eisenberg, E., and Greene, L.E., Expression of auxilin or AP180 inhibits endocytosis by mislocalizing clathrin: evidence for formation of nascent pits containing AP1 or AP2 but not clathrin, J. Cell Sci., 2001, vol. 114, p. 353.
Wang, X.J., Luo, Y.P., Shi, L., Pang, P.X., and Gao, G., Analysis of expression characteristics of scarecrow-like gene Stsl-1 elicited by exogenous hormone and Ralstonia solanacearum infection in potato, Int. J. Agric. Biol., 2019, vol. 22, p. 201.
Kanokporn, B., John, C. S., Rujin, C., Simon, G., and Patrick, H. M., Altered response to gravity is a peripheral membrane protein that modulates gravity-induced cytoplasmic alkalinization and lateral auxin transport in plant statocytes, Plant Cell, 2003, vol. 15, p. 2612.
Yang, K.Z., Xia, C., Liu, X.L., Dou, X.Y., Wang, W., Chen, L.Q., Zhang, X.Q., Xie, L.F., He, L., and Ma, X., A mutation in Thermosensitive Male Sterile 1, encoding a heat shock protein with DnaJ and PDI domains, leads to thermosensitive gametophytic male sterility in Arabidopsis, Plant J., 2009, vol. 57, p. 870.
Sun, J.X., Xiao, T.T., Nie, J.T., Chen, Y., Lv, D., Pan, M., Gao, Q.F., Guo, C.L., Zhang, L.Y., He, H.L., Lian, H.L., Pan, J.S., Cai, R., and Wang, G., Mapping and identification of CsUp, a gene encoding an auxilin-like protein, as a putative candidate gene for the upwardpedicel mutation (up) in cucumber, BMC Plant Biol., 2019, vol. 19, p. 157.
ACKNOWLEDGMENTS
We would like to thank the institutes of Vegetables and Flowers and Plant Protection of the Chinese Academy of Agricultural Sciences for potato materials.
Funding
This study was supported by the National Natural Science Foundation of China (project nos. 31771858 and 31271774).
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Shi, L., Luo, Y.P., Wang, X.J. et al. Molecular Cloning and Expression Analysis of Auxilin-Like Gene StAL1 in Potato (Solanum tuberosum). Russ J Plant Physiol 68, 56–65 (2021). https://doi.org/10.1134/S1021443721010167
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DOI: https://doi.org/10.1134/S1021443721010167