Proteomic dataset of knockdown of AKT3 on protein expression and function in female germline stem cells
Yue Shen1, Chunlan Mu1, Qingling Jia1
1Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China.
Abstract:
Female germline stem cells (FGSCs) are adult stem cells capable of self-renewal and differentiation into mature oocytes. AKT3, a member of the AKT kinase family, plays crucial roles in multiple cellular processes, such as proliferation, migration, and apoptosis. However, the mechanism by which AKT3 affects the development of FGSCs is poorly understood. We performed 4D-data-independent acquisition (DIA) Quantitative Proteomics on mouse FGSCs in which AKT3 was knocked down using a lentivirus and on control FGSCs. Based on the raw DIA data, coupled with database searches and data filtering, we identified 46,260 peptides, including 45,821 unique peptides, corresponding to 6849 identified proteins and 6697 comparable proteins. These identified proteins were functionally annotated using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Protein Domain, Clusters of Orthologous Genes (COG)/Eukaryotic Orthologous Groups (KOG), STRING database, Reactome, WikiPathways, HallMark, and transcription factor (TF) analyses. Fisher's exact test was used to assess the significance of functional enrichment of the differentially abundant proteins. We identified 281 differentially abundant proteins between AKT3 knockdown and control FGSCs, comprising 229 upregulated and 52 downregulated proteins. We performed clustering analysis on these differentially abundant proteins based on functional enrichment using GO, Domain, KEGG, Reactome and WikiPathways platforms. A protein-protein interaction network was constructed to demonstrate interactions between proteins. These datasets will facilitate future investigations into the mechanisms governing FGSC self-renewal and differentiation and will provide a foundation for understanding diseases related to abnormal germ cell development.
Insights
This study reveals how AKT3 influences female germline stem cell development by identifying 281 differentially abundant proteins. These findings offer new insights into FGSC self-renewal, differentiation, and related diseases.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Proteomics
Background:
- Female germline stem cells (FGSCs) are vital for oocyte production, but the regulatory mechanisms are not fully understood.
- AKT3 is implicated in cell proliferation and apoptosis, yet its specific role in FGSC development remains unclear.
Purpose of the Study:
- To investigate the proteomic changes in mouse FGSCs upon AKT3 knockdown.
- To identify proteins and pathways affected by AKT3, providing insights into FGSC self-renewal and differentiation.
Main Methods:
- Utilized 4D-data-independent acquisition (DIA) quantitative proteomics on mouse FGSCs with and without AKT3 knockdown.
- Identified and quantified thousands of peptides and proteins, followed by extensive functional annotation and pathway analysis.
- Constructed a protein-protein interaction network to visualize molecular relationships.
Main Results:
- Identified 6,697 comparable proteins between control and AKT3-knockdown FGSCs.
- Discovered 281 differentially abundant proteins, with 229 upregulated and 52 downregulated in AKT3-knockdown cells.
- Functional enrichment analysis revealed significant alterations in key cellular processes.
Conclusions:
- The proteomic landscape of FGSCs is significantly altered by AKT3 knockdown.
- These findings provide a foundation for understanding the molecular mechanisms of FGSC self-renewal, differentiation, and germ cell development disorders.
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