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Previously, we showed that full-length RAG1 stability is regulated by viral protein R binding protein (VprBP) through its association with an amino-terminal region in RAG1, but the mechanism remains unclear. As an unbiased approach to uncover potential cofactors involved in the process, we compared protein interactomes between RAG1\/RAG2 complexes formed when the amino-terminal third of RAG1 was present or absent. These experiments identified RACK1 as preferentially associating with full-length RAG1. Because RACK1 is implicated in mediating protein degradation in other contexts, we evaluated how loss of RACK1 in B cells affects B cell development and V(D)J recombination. We find that conditional disruption of Rack1 expression in the B lineage in mice blocks B cell development at the pro\u2013B cell stage and impairs V(D)J recombination after Igh DH-JH rearrangement. In this background, enforced Bcl2 expression does not significantly rescue B cell development but does enable the V(D)J recombination defect to be bypassed. However, the phenotype of these mice does not show the excessive Igk rearrangement, skewing toward Ig\u03bb+ B cells, or increased RAG1 protein levels observed when VprBP expression is similarly disrupted in B cells, arguing against RACK1 serving as a cofactor in RAG1 degradation. Further studies provide evidence that loss of RACK1 in primary B cells dysregulates cell cycle progression, apoptosis, proliferation, and signaling through MAPK and NF-\u03baB pathways.<\/jats:p>","DOI":"10.1093\/jimmun\/vkaf217","type":"journal-article","created":{"date-parts":[[2025,8,24]],"date-time":"2025-08-24T15:58:19Z","timestamp":1756051099000},"page":"3003-3016","source":"Crossref","is-referenced-by-count":1,"title":["RACK1 is required for normal B cell development and signaling but not RAG1 degradation"],"prefix":"10.1093","volume":"214","author":[{"given":"Victoria L","family":"Palmer","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology and Immunology, Creighton University , Omaha, NE,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"N Max","family":"Schabla","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Immunology, Creighton University , Omaha, NE,","place":["United 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