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Identification of Key Amino Acid Residues Involved in the Localization of Sorting Nexin 10 and Induction of Vacuole Formation

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Abstract

Sorting nexin 10 (SNX10) induces formation of vacuoles participating in the endosome morphogenesis in mammalian cells, but the key amino acids involved in this function have not been fully identified. In this study, point mutations were introduced to the conserved region of the SNX10 PX domain to elucidate the function of these key amino acid residues. The number of vacuoles in the R53A mutant was partially decreased, while the R52A and R51A mutants completely lacked the vacuoles. All mutant proteins lost the phosphatidylinositol 3-phosphate (PtdIns3P)-binding ability and endosomal localization. Retargeting the mutants to the endosomes rescued partially or fully the vacuole-inducing ability in the R51A and R53A mutants, respectively, but not in the R52A mutant. No vacuoles were induced when the R51A mutant was targeted to other organelles. Structural analysis showed that Arg53 is responsible for the PtdIns(3)P binding, whereas Arg51 and Arg52 contribute to the structural integrity of SNX10. We conclude that the disruption of the key residues affects the structure and function of SNX10 and that induction of vacuole formation by SNX10 depends on its endosomal location.

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Abbreviations

COX8:

cytochrome c oxidase subunit VIII

JMJD6:

jumonji domain containing 6

MTS:

mitochondrial target sequence

NLS:

nuclear location sequence

PX:

Phox homology

SNX10:

sorting nexin 10

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Funding

This study was supported by the National Natural Science Foundation of China (81560005 and 81760007), Natural Science Foundation of Guangxi province (project no. 2018GXNSFAA138052), and The Guangxi Medical Care Appropriate Technology Research and Development (project no. S201603).

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Authors and Affiliations

  1. The Laboratory of Respiratory Disease, The Affiliated Hospital of Guilin Medical University, 541000, Guilin, China

    Bo Xiao, Lixia Hou & Dong Yao

  2. Department of Basic Medical Sciences, School of Nursing, Sanmenxia Polytechnic, 472000, Sanmenxia, China

    Nana Liu

  3. Department of Respiratory and Critical Care Medicine, The Affiliated Hospital of Guilin Medical University, 541000, Guilin, China

    Ming Jiang & Dong Yao

  4. The Key Laboratory of Respiratory Disease, Education Department of Guangxi Zhuang Autonomous Region, 541000, Guilin, China

    Dong Yao

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  1. Bo Xiao
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  2. Nana Liu
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  3. Lixia Hou
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  4. Ming Jiang
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  5. Dong Yao
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Correspondence to Dong Yao.

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The authors declare no conflicts of interest. This article does not contain any studies involving human participants or animals performed by any of the authors.

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Xiao, B., Liu, N., Hou, L. et al. Identification of Key Amino Acid Residues Involved in the Localization of Sorting Nexin 10 and Induction of Vacuole Formation. Biochemistry Moscow 86, 1377–1387 (2021). https://doi.org/10.1134/S000629792111002X

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  • DOI: https://doi.org/10.1134/S000629792111002X

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