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Title: Overexpression of Rice Auxilin-Like Protein, XB21, Induces Necrotic Lesions, up-Regulates Endocytosis-Related Genes, and Confers Enhanced Resistance to Xanthomonas oryzae pv. oryzae

Abstract

The rice immune receptor XA21 confers resistance to the bacterial pathogen, Xanthomonas oryzae pv. oryzae (Xoo). To elucidate the mechanism of XA21-mediated immunity, we previously performed a yeast two-hybrid screening for XA21 interactors and identified XA21 binding protein 21 (XB21). Here, we report that XB21 is an auxilin-like protein predicted to function in clathrin-mediated endocytosis. We demonstrate an XA21/XB21 in vivo interaction using co-immunoprecipitation in rice. Overexpression of XB21 in rice variety Kitaake and a Kitaake transgenic line expressing XA21 confers a necrotic lesion phenotype and enhances resistance to Xoo. RNA sequencing reveals that XB21 overexpression results in the differential expression of 8735 genes (4939 genes up- and 3846 genes down-regulated) (≥2-folds, FDR ≤0.01). The up-regulated genes include those predicted to be involved in ‘cell death’ and ‘vesicle-mediated transport’. These results indicate that XB21 plays a role in the plant immune response and in regulation of cell death. The up-regulation of genes controlling ‘vesicle-mediated transport’ in XB21 overexpression lines is consistent with a functional role for XB21 as an auxilin.

Authors:
 [1];  [2];  [3];  [2]
  1. Univ. of California, Davis, CA (United States). Dept. of Plant Pathology and the Genome Center; Sejong Univ., Seoul (Korea, Republic of). Dept. of Bioresources Engineering and the Plant Engineering Research Inst.
  2. Univ. of California, Davis, CA (United States). Dept. of Plant Pathology and the Genome Center; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst., Feedstocks Division; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology
  3. Univ. of California, Davis, CA (United States). Dept. of Plant Pathology and the Genome Center; Jawaharlal Nehru Univ., New Delhi (India). School of Computational & Integrative Sciences
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Science Foundation (NSF); National Institutes of Health (NIH)
OSTI Identifier:
1379882
Grant/Contract Number:
AC02-05CH11231; IOS-0817738; GM55962
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Rice
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 1939-8425
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Auxilin; Auxilin-Like Protein; Disease Resistance; Heat Shock Cognate Protein; Heat Shock Protein; XA21 Immune Receptor; XA21 Binding Protein 21; Xanthomonas oryzae pv. oryzae

Citation Formats

Park, Chang-Jin, Wei, Tong, Sharma, Rita, and Ronald, Pamela 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. United States: N. p., 2017. Web. doi:10.1186/s12284-017-0166-1.
Park, Chang-Jin, Wei, Tong, Sharma, Rita, & Ronald, Pamela 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. United States. doi:10.1186/s12284-017-0166-1.
Park, Chang-Jin, Wei, Tong, Sharma, Rita, and Ronald, Pamela C. 2017. "Overexpression of Rice Auxilin-Like Protein, XB21, Induces Necrotic Lesions, up-Regulates Endocytosis-Related Genes, and Confers Enhanced Resistance to Xanthomonas oryzae pv. oryzae". United States. doi:10.1186/s12284-017-0166-1. https://www.osti.gov/servlets/purl/1379882.
@article{osti_1379882,
title = {Overexpression of Rice Auxilin-Like Protein, XB21, Induces Necrotic Lesions, up-Regulates Endocytosis-Related Genes, and Confers Enhanced Resistance to Xanthomonas oryzae pv. oryzae},
author = {Park, Chang-Jin and Wei, Tong and Sharma, Rita and Ronald, Pamela C.},
abstractNote = {The rice immune receptor XA21 confers resistance to the bacterial pathogen, Xanthomonas oryzae pv. oryzae (Xoo). To elucidate the mechanism of XA21-mediated immunity, we previously performed a yeast two-hybrid screening for XA21 interactors and identified XA21 binding protein 21 (XB21). Here, we report that XB21 is an auxilin-like protein predicted to function in clathrin-mediated endocytosis. We demonstrate an XA21/XB21 in vivo interaction using co-immunoprecipitation in rice. Overexpression of XB21 in rice variety Kitaake and a Kitaake transgenic line expressing XA21 confers a necrotic lesion phenotype and enhances resistance to Xoo. RNA sequencing reveals that XB21 overexpression results in the differential expression of 8735 genes (4939 genes up- and 3846 genes down-regulated) (≥2-folds, FDR ≤0.01). The up-regulated genes include those predicted to be involved in ‘cell death’ and ‘vesicle-mediated transport’. These results indicate that XB21 plays a role in the plant immune response and in regulation of cell death. The up-regulation of genes controlling ‘vesicle-mediated transport’ in XB21 overexpression lines is consistent with a functional role for XB21 as an auxilin.},
doi = {10.1186/s12284-017-0166-1},
journal = {Rice},
number = 1,
volume = 10,
place = {United States},
year = 2017,
month = 6
}

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  • Background: The rice receptor kinase XA21 confers robust resistance to the bacterial pathogen Xanthomonas oryzae pv. oryzae( Xoo). We previously reported that XA21 is cleaved in transgenic plants overexpressing XA21 with a GFP tag ( Ubi-XA21-GFP) and that the released C-terminal domain is localized to the nucleus. XA21 carries a predicted nuclear localization sequence (NLS) that directs the C-terminal domain to the nucleus in transient assays, whereas alanine substitutions in the NLS disrupt the nuclear localization. Methods: To determine if the predicted NLS is required for XA21-mediated immunity in planta, we generated transgenic plants overexpressing an XA21 variant carrying themore » NLS with the same alanine substitutions ( Ubi-XA21nls-GFP). Results: Ubi- XA21nls-GFP plants displayed slightly longer lesion lengths, higher Xoo bacterial populations after inoculation and lower levels of reactive oxygen species production compared with the Ubi- XA21-GFP control plants. However, the Ubi- XA21nls-GFP plants express lower levels of protein than that observed in Ubi- XA21-GFP. Discussion: These results demonstrate that the predicted NLS is not required for XA21-mediated immunity.« less
  • XometC, a cystathionine γ-lyase-like protein from X. oryzae pv. oryzae and an antibacterial drug-target protein against bacterial blight, was cloned, purified and crystallized. Preliminary X-ray crystallographic analysis of XometC crystals was carried out. Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight of rice (Oryza sativa L.), one of the most devastating diseases of rice in most rice-growing countries. XometC, a cystathionine γ-lyase (CGL) like protein that is an antibacterial drug-target protein against Xoo, was cloned, expressed, purified and crystallized. CGL catalyzes the second step in the reverse-transsulfuration pathway, which is essential for the metabolic interconversion of the sulfur-containing amino acidsmore » cysteine and methionine. Crystals of two different shapes, plate-shaped and pyramid-shaped, diffracted to 2.9 and 3.2 Å resolution and belonged to the primitive orthogonal space group P2{sub 1}2{sub 1}2{sub 1} and the tetragonal space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = 73.0, b = 144.9, c = 152.3 Å and a = b = 78.2, c = 300.7 Å, respectively. For the P2{sub 1}2{sub 1}2{sub 1} crystals, three or four monomers exist in the asymmetric unit with a corresponding V{sub M} of 3.02 or 2.26 Å{sup 3} Da{sup −1} and a solvent content of 59.3 or 45.7%. For the P4{sub 1} (or P4{sub 3}) crystals, four or five monomers exist in the asymmetric unit with a corresponding V{sub M} of 2.59 or 2.09 Å{sup 3} Da{sup −1} and a solvent content of 52.5 or 40.6%.« less
  • The nuclear location and relocation of genes play crucial regulatory roles in gene expression. SATB1, a MAR-binding protein, has been found to regulate {beta}-like globin genes through chromatin remodeling. In this study, we generated K562 cells over-expressing wild-type or nuclear matrix targeting sequences (NMTS)-deficient SATB1 and found that like wild-type SATB1, NMTS-deficient SATB1 induces out loop of {beta}-globin cluster from its chromosome territory (CT), while it is unable to associate the cluster with the nuclear matrix as wild-type SATB1 does and had no regulatory functions to the {beta}-globin cluster. Besides, our data showed that the transacting factor occupancies and chromatinmore » modifications at {beta}-globin cluster were differentially affected by wild-type and NMTS-deficient SATB1. These results indicate that SATB1 regulates {beta}-like globin genes at the nuclear level interlaced with chromatin and DNA level, and emphasize the nuclear matrix binding activity of SATB1 to its regulatory function.« less