Identification of novel interacting partners of the NEDD4 ubiquitin ligase in mouse testis.

Autor: Manning J; Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA 5001, Australia. Electronic address: jantina.manning@unisa.edu.au., Windley SP; Department of Anatomy and Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia., Sandow JJ; The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia., Shah SS; Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA 5001, Australia., Western P; Centre for Reproductive Health, Hudson Institute of Medical Research, Clayton, VIC 3800, Australia., Wilhelm D; Department of Anatomy and Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia., Kumar S; Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA 5001, Australia. Electronic address: sharad.kumar@unisa.edu.au.
Jazyk: angličtina
Zdroj: Journal of proteomics [J Proteomics] 2020 Jul 15; Vol. 223, pp. 103830. Date of Electronic Publication: 2020 May 22.
DOI: 10.1016/j.jprot.2020.103830
Abstrakt: Posttranslational modification by ubiquitination targets proteins for degradation, recycling, stabilization or altered trafficking, and as such can alter cellular signaling pathways. The substrate specificity of this multistep process is controlled by ubiquitin ligases, including those of the HECT domain-containing NEDD4 family. In the testis, ubiquitination of many proteins contributes to organ development and maturation of spermatozoa and NEDD4 is known to be important in the control of spermatogonial stem cell homeostasis. However, a comprehensive understanding of NEDD4 substrates in testis development is lacking. Here we demonstrate high expression of Nedd4 in somatic cells of the mouse testis and in the murine Leydig cell-like cell line TM3. Immunoprecipitation of NEDD4 tagged with GFP at either the amino or carboxyl terminus was subjected to proteomic analysis for interacting proteins. We identified a substantial list of potential interaction partners, including known NEDD4 substrates, proteins involved in ubiquitination and proteins important for testis development and spermatogenesis. We confirmed the interaction of NEDD4 with a subset of these putative interacting proteins, validating the integrity of the dataset. These potential interactors may be further explored to reveal important roles of NEDD4-mediated ubiquitination in the testis. SIGNIFICANCE: Ubiquitination is important for testis development and function, and NEDD4 is known to ubiquitinate various proteins to affect cellular signaling and development, including those implicated in spermatogenesis. However, substrates of NEDD4 that are important during testis development remain to be identified. Here we report NEDD4 expression in the developing testis and TM3 testicular cell line. This study identifies a substantial list of NEDD4 interacting proteins in the TM3 testicular cell line, with validation of some of these interactions. Hence, this provides novel NEDD4 targets that may contribute to testis development and function that may be further explored.
Competing Interests: Declaration of Competing Interest The authors declare that no conflicts of interest exist.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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