An in vivo “turning model” reveals new RanBP9 interactions in lung macrophages
The biological functions of the scaffold protein Ran Binding Protein 9 (RanBP9) remain elusive in macrophages or any other cell type where this protein is expressed together with its CTLH (C-terminal to LisH) complex partners. We have engineered a new mouse model, named RanBP9-TurnX, where RanBP9 fused to three copies of the HA tag (RanBP9-3xHA) can be turned into RanBP9-V5 tagged upon Cre-mediated recombination. We created this model to enable stringent biochemical studies at cell type specific level throughout the entire organism. Here, we have used this tool crossed with LysM-Cre transgenic mice to identify RanBP9 interactions in lung macrophages. We show that RanBP9-V5 and RanBP9-3xHA can be both co-immunoprecipitated with the known members of the CTLH complex from the same whole lung lysates. However, more than ninety percent of the proteins pulled down by RanBP9-V5 differ from those pulled-down by RanBP9-HA. The lung RanBP9-V5 associated proteome includes previously unknown interactions with macrophage-specific proteins as well as with players of the innate immune response, DNA damage response, metabolism, and mitochondrial function. This work provides the first lung specific RanBP9-associated interactome in physiological conditions and reveals that RanBP9 and the CTLH complex could be key regulators of macrophage bioenergetics and immune functions.
Authors
- Lino Tessarollo (ORCID: https://orcid.org/0000-0001-6420-772X)
- Anna Tessari (ORCID: https://orcid.org/0000-0002-6036-1954)
- Amer Ahmed (ORCID: https://orcid.org/0000-0002-0503-4045)
- Li‐wen Zhang (ORCID: https://orcid.org/0000-0002-3951-5031)
- Yasuko Kajimura (ORCID: https://orcid.org/0000-0002-0819-6904)
- Piero Del Boccio (ORCID: https://orcid.org/0000-0003-1653-2194)
- Vincenzo Coppola (ORCID: https://orcid.org/0000-0001-6163-1779)
- Federica Di Marco (ORCID: https://orcid.org/0000-0002-1281-607X)
- Arturo Orlacchio (ORCID: https://orcid.org/0000-0002-6044-4258)
- Lara Rizzotto (ORCID: https://orcid.org/0000-0002-2658-9103)
- David P. Carbone (ORCID: https://orcid.org/0000-0003-3002-1921)
- Giuseppe Fiermonte (ORCID: https://orcid.org/0000-0002-6764-9395)
- Shimaa Soliman (ORCID: https://orcid.org/0000-0001-6517-9847)
- Maria Concetta Cufaro (ORCID: https://orcid.org/0000-0002-6457-7795)
- Dario Palmieri (ORCID: https://orcid.org/0000-0002-0797-4268)
- Foued Amari (ORCID: https://orcid.org/0000-0003-4904-877X)
- Damu Sunilkumar (ORCID: https://orcid.org/0000-0003-0376-6039)
- Mike Freitas
- Min Chen (ORCID: https://orcid.org/0000-0001-8033-9116)
- Alessia Lodi (ORCID: https://orcid.org/0000-0002-2104-5713)
- Alexandra Thoms
- Shuxin Dong
- Joseph M. Amann
Institutions
- Northwestern University (US)
- Kenyon College (US)
- The Ohio State University Comprehensive Cancer Center – Arthur G. James Cancer Hospital and Richard J. Solove Research Institute (US)
- The Ohio State University Wexner Medical Center (US)
- University of Chieti-Pescara (IT)
- Yamaguchi University Hospital (JP)
- Center for Cancer Research (US)
- University of Bari Aldo Moro (IT)
- The University of Texas at Austin (US)
Publication Details
- Journal
- Cell Death Discovery
- Published
- 2025-04-13
- DOI
- https://doi.org/10.1038/s41420-025-02456-2
- Citations
- 4
- Primary Topic
- Ubiquitin and proteasome pathways
- Type
- article
- Field-Weighted Citation Impact
- 3.22
Funders
- Ohio State University
- Pelotonia
- National Institutes of Health
- National Cancer Institute