CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line

Abstract Background Human spermatogonial stem cells (SSCs) exhibit a remarkable capacity for proliferation, crucial for sustaining spermatogenesis throughout life. While the Cullin-RING E3 ubiquitin ligase 2 (CRL2) complex is known to regulate various cellular functions, its precise role in human SS...

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Main Authors: Bing Jiang, Kehan Wang, Haoyue Hu, Wenxin Gao, Cong Shen, Xia Chen, Xiaoyan Huang, Jun Yu, Yibo Wu, Bo Zheng
Format: Article
Language:English
Published: BMC 2025-08-01
Series:BMC Biology
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Online Access:https://doi.org/10.1186/s12915-025-02363-z
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author Bing Jiang
Kehan Wang
Haoyue Hu
Wenxin Gao
Cong Shen
Xia Chen
Xiaoyan Huang
Jun Yu
Yibo Wu
Bo Zheng
author_facet Bing Jiang
Kehan Wang
Haoyue Hu
Wenxin Gao
Cong Shen
Xia Chen
Xiaoyan Huang
Jun Yu
Yibo Wu
Bo Zheng
author_sort Bing Jiang
collection DOAJ
description Abstract Background Human spermatogonial stem cells (SSCs) exhibit a remarkable capacity for proliferation, crucial for sustaining spermatogenesis throughout life. While the Cullin-RING E3 ubiquitin ligase 2 (CRL2) complex is known to regulate various cellular functions, its precise role in human SSCs has not been fully elucidated. This study aimed to investigate a novel variant of the CRL2 complex, termed CRL2LRRC41, and its role in SSC function. Methods We utilized molecular biology techniques, including gene knockdown and functional assays, to assess the effects of CRL2LRRC41 on the proliferative and migratory abilities of human spermatogonial stem cell-like cell (SSCLC) line. Additionally, we employed proteomics and biochemical approaches to identify potential substrates of CRL2LRRC41. We specifically focused on ATP-dependent RNA helicase DDX5, a known regulator of spermatogenesis, to explore its interaction with CRL2LRRC41 and the downstream molecular mechanisms involved. Results Our findings revealed that the disruption or dysfunction of CRL2LRRC41 led to reduced proliferative and migratory abilities in human SSCLCs. Through our investigation, we identified DDX5 as a ubiquitination substrate of CRL2LRRC41. Notably, the ubiquitination of DDX5 fosters its interaction with the RNA-binding protein ELAVL1, without directing DDX5 towards degradation via the ubiquitin–proteasome system (UPS). This interaction enhances the stability of the downstream transcript, Noggin (NOG), thereby supporting human SSCLC proliferation and migration. Conclusions This study provides the first identification of the CRL2LRRC41 complex in human SSCLCs and elucidates the molecular mechanisms by which CRL2LRRC41 facilitates SSCLC function via ubiquitination-mediated protein interactions. These findings offer novel insights into the molecular underpinnings of male infertility.
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spelling doaj-art-83c3c2eedb174b4e8a39373d9be4fd0f2025-08-20T03:41:57ZengBMCBMC Biology1741-70072025-08-0123111910.1186/s12915-025-02363-zCRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell LineBing Jiang0Kehan Wang1Haoyue Hu2Wenxin Gao3Cong Shen4Xia Chen5Xiaoyan Huang6Jun Yu7Yibo Wu8Bo Zheng9Human Reproductive and Genetic Center, Affiliated Hospital of Jiangnan UniversityState Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproduction and Genetics, Gusu School, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Nanjing Medical UniversityHuman Reproductive and Genetic Center, Affiliated Hospital of Jiangnan UniversityState Key Laboratory of Reproductive Medicine and Offspring Health, Department of Histology and Embryology, School of Basic Medical Sciences, Nanjing Medical UniversityState Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproduction and Genetics, Gusu School, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Nanjing Medical UniversityCenter of Reproductive Medicine, Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong UniversityState Key Laboratory of Reproductive Medicine and Offspring Health, Department of Histology and Embryology, School of Basic Medical Sciences, Nanjing Medical UniversityInstitute of Reproductive Medicine, Medical School of Nantong University, Nantong UniversityHuman Reproductive and Genetic Center, Affiliated Hospital of Jiangnan UniversityState Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproduction and Genetics, Gusu School, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Nanjing Medical UniversityAbstract Background Human spermatogonial stem cells (SSCs) exhibit a remarkable capacity for proliferation, crucial for sustaining spermatogenesis throughout life. While the Cullin-RING E3 ubiquitin ligase 2 (CRL2) complex is known to regulate various cellular functions, its precise role in human SSCs has not been fully elucidated. This study aimed to investigate a novel variant of the CRL2 complex, termed CRL2LRRC41, and its role in SSC function. Methods We utilized molecular biology techniques, including gene knockdown and functional assays, to assess the effects of CRL2LRRC41 on the proliferative and migratory abilities of human spermatogonial stem cell-like cell (SSCLC) line. Additionally, we employed proteomics and biochemical approaches to identify potential substrates of CRL2LRRC41. We specifically focused on ATP-dependent RNA helicase DDX5, a known regulator of spermatogenesis, to explore its interaction with CRL2LRRC41 and the downstream molecular mechanisms involved. Results Our findings revealed that the disruption or dysfunction of CRL2LRRC41 led to reduced proliferative and migratory abilities in human SSCLCs. Through our investigation, we identified DDX5 as a ubiquitination substrate of CRL2LRRC41. Notably, the ubiquitination of DDX5 fosters its interaction with the RNA-binding protein ELAVL1, without directing DDX5 towards degradation via the ubiquitin–proteasome system (UPS). This interaction enhances the stability of the downstream transcript, Noggin (NOG), thereby supporting human SSCLC proliferation and migration. Conclusions This study provides the first identification of the CRL2LRRC41 complex in human SSCLCs and elucidates the molecular mechanisms by which CRL2LRRC41 facilitates SSCLC function via ubiquitination-mediated protein interactions. These findings offer novel insights into the molecular underpinnings of male infertility.https://doi.org/10.1186/s12915-025-02363-zLRRC41DDX5ELAVL1UbiquitinationSpermatogonial Stem Cells
spellingShingle Bing Jiang
Kehan Wang
Haoyue Hu
Wenxin Gao
Cong Shen
Xia Chen
Xiaoyan Huang
Jun Yu
Yibo Wu
Bo Zheng
CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
BMC Biology
LRRC41
DDX5
ELAVL1
Ubiquitination
Spermatogonial Stem Cells
title CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
title_full CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
title_fullStr CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
title_full_unstemmed CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
title_short CRL2LRRC41-Mediated DDX5 Ubiquitination Enhances Interaction with ELAVL1 Preventing NOG mRNA Degradation and Sustaining Proliferation and Migration of Human Spermatogonial Stem Cell-Like Cell Line
title_sort crl2lrrc41 mediated ddx5 ubiquitination enhances interaction with elavl1 preventing nog mrna degradation and sustaining proliferation and migration of human spermatogonial stem cell like cell line
topic LRRC41
DDX5
ELAVL1
Ubiquitination
Spermatogonial Stem Cells
url https://doi.org/10.1186/s12915-025-02363-z
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