Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Rack1 regulates B-cell development and function by binding to and stabilizing the transcription factor Pax5

Cellular & Molecular Immunology (2024)Cite this article

Subjects

Abstract

The transcription factor Pax5 activates genes essential for B-cell development and function. However, the regulation of Pax5 expression remains elusive. The adaptor Rack1 can interact with multiple transcription factors and modulate their activation and/or stability. However, its role in the transcriptional control of B-cell fates is largely unknown. Here, we show that CD19-driven Rack1 deficiency leads to pro-B accumulation and a simultaneous reduction in B cells at later developmental stages. The generation of bone marrow chimeras indicates a cell-intrinsic role of Rack1 in B-cell homeostasis. Moreover, Rack1 augments BCR and TLR signaling in mature B cells. On the basis of the aberrant expression of Pax5-regulated genes, including CD19, upon Rack1 deficiency, further exploration revealed that Rack1 maintains the protein level of Pax5 through direct interaction and consequently prevents Pax5 ubiquitination. Accordingly, Mb1-driven Rack1 deficiency almost completely blocks B-cell development at the pro-B-cell stage. Ectopic expression of Pax5 in Rack1-deficient pro-B cells partially rescues B-cell development. Thus, Rack1 regulates B-cell development and function through, at least partially, binding to and stabilizing Pax5.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Ripperger TJ, Bhattacharya D. Transcriptional and metabolic control of memory B cells and plasma cells. Annu Rev Immunol. 2021;39:345–68.

    Article  PubMed  CAS  Google Scholar 

  2. Bassing CH, Swat W, Alt FW. The mechanism and regulation of chromosomal V(D)J recombination. Cell. 2002;109:S45–55.

    Article  PubMed  CAS  Google Scholar 

  3. Berland R, Wortis HH. Origins and functions of B-1 cells with notes on the role of CD5. Annu Rev Immunol. 2002;20:253–300.

    Article  PubMed  CAS  Google Scholar 

  4. Cerutti A, Cols M, Puga I. Marginal zone B cells: virtues of innate-like antibody-producing lymphocytes. Nat Rev Immunol. 2013;13:118–32.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Victora GD, Nussenzweig MC. Germinal centers. Annu Rev Immunol. 2012;30:429–57.

    Article  PubMed  CAS  Google Scholar 

  6. Nutt SL, Hodgkin PD, Tarlinton DM, Corcoran LM. The generation of antibody-secreting plasma cells. Nat Rev Immunol. 2015;15:160–71.

    Article  PubMed  CAS  Google Scholar 

  7. Nutt SL, Heavy B, Rolink AG, Busslinger M. Commitment to the B-lymphoid lineage depends on the transcription factor Pax5. Nature. 1999;401:556–62.

    Article  PubMed  CAS  Google Scholar 

  8. Horcher M, Souabni A, Busslinger M. Pax5/BSAP maintains the identity of B cells in late B lymphopoiesis. Immunity. 2001;14:779–90.

    Article  PubMed  CAS  Google Scholar 

  9. Calderón L, Schindler K, Malin SG, Schebesta A, Sun Q, Schwickert T, et al. Pax5 regulates B-cell immunity by promoting PI3K signaling via PTEN downregulation. Sci Immunol. 2021;6:eabg5003.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Kaiser FMP, Gruenbacher S, Oyaga MR, Nio E, Jaritz M, Sun Q, et al. Biallelic PAX5 mutations cause hypogammaglobulinemia, sensorimotor deficits, and autism spectrum disorder. J Exp Med. 2022;219:e20220498.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Fuxa M, Busslinger M. Reporter gene insertions reveal a strictly B lymphoid-specific expression pattern of Pax5 in support of its B-cell identity function. J Immunol. 2007;178:3031–7.

    Article  PubMed  CAS  Google Scholar 

  12. Delogu A, Schebesta A, Sun Q, Aschenbrenner K, Perlot T, Busslinger M. Gene repression by Pax5 in B cells is essential for blood cell homeostasis and is reversed in plasma cells. Immunity. 2006;24:269–81.

    Article  PubMed  CAS  Google Scholar 

  13. Revilla-i-Domingo R, Bilic I, Vilagos B, Tahoh H, Ebert A, Tamir IM, et al. The B-cell identity factor Pax5 regulates distinct transcriptional programs in early and late B lymphopoiesis. EMBO J. 2012;31:3130–46.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Schebesta A, McManus S, Salvagiotto G, Delogu A, Busslinger GA, Busslinger M. Transcription factor Pax5 activates the chromatin of key genes involved in B-cell signaling, adhesion, migration, and immune function. Immunity. 2007;27:49–63.

    Article  PubMed  CAS  Google Scholar 

  15. Nutt SL, Morrison AM, Dörfler P, Rolink A, Busslinger M. Identification of BSAP (Pax-5) target genes in early B-cell development by loss- and gain-of-function experiments. EMBO J. 1998;17:2319–33.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Pasqualucci L, Neumeister P, Goossens T, Nanjangud G, Chaganti RS, Küppers R, Dalla-Favera R. Hypermutation of multiple proto-oncogenes in B-cell diffuse large-cell lymphomas. Nature. 2001;412:341–6.

    Article  PubMed  CAS  Google Scholar 

  17. Busslinger M, Klix N, Pfeffer P, Graninger PG, Kozmik Z. Deregulation of PAX-5 by translocation of the Eµ enhancer of the IgH locus adjacent to two alternative PAX-5 promoters in a diffuse large-cell lymphoma. Proc Natl Acad Sci USA. 1996;93:6129–34.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Lida S, Rao PH, Nallasivam P, Hibshoosh H, Butler M, Louie DC, et al. The t(9;14)(p13;q32) chromosomal translocation associated with lymphoplasmacytoid lymphoma involves the PAX-5 gene. Blood. 1996;88:4110–7.

    Article  CAS  Google Scholar 

  19. Morrison AM, Jäger U, Chott A, Schebesta M, Haas OA, Busslinger M. Deregulated PAX-5 transcription from a translocated IgH promoter in marginal zone lymphoma. Blood. 1998;92:3865–78.

    Article  PubMed  CAS  Google Scholar 

  20. Cozma D, Yu D, Hodawadekar S, Azvolinsky A, Grande S, Tobias JW, et al. B-cell activator PAX5 promotes lymphomagenesis through stimulation of B-cell receptor signaling. J Clin Invest. 2007;117:2602–10.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Usacheva A, Smith R, Minshall R, Baida G, Seng S, Croze E, et al. The WD motif-containing protein receptor for activated protein kinase C (RACK1) is required for recruitment and activation of signal transducer and activator of transcription 1 through the type I interferon receptor. J Biol Chem. 2001;276:22948–53.

    Article  PubMed  CAS  Google Scholar 

  22. Zhang W, Zong CS, Hermanto U, Lopez-Bergami P, Ronai Z, Wang LH. RACK1 recruits STAT3 specifically to insulin and insulin-like growth factor 1 receptors for activation, which is important for regulating anchorage-independent growth. Mol Cell Biol. 2006;26:413–24.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Okano K, Schnaper HW, Bomsztyk K, Hayashida T. RACK1 binds to Smad3 to modulate transforming growth factor-β-stimulated α2(I) collagen transcription in renal tubular epithelial cells. J Biol Chem. 2006;281:26196–204.

    Article  PubMed  CAS  Google Scholar 

  24. Zhu Q, Chen L, Li Y, Huang M, Shao J, Li S, et al. Rack1 is essential for corticogenesis by preventing p21-dependent senescence in neural stem cells. Cell Rep. 2021;36:109639.

    Article  PubMed  CAS  Google Scholar 

  25. Liu YV, Baek JH, Zhang H, Diez R, Cole RN, Semenza GL. RACK1 competes with HSP90 for binding to HIF-1α and is required for O2-independent and HSP90 inhibitor-induced degradation of HIF-1α. Mol Cell. 2007;25:207–17.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Wang J, Chen X, Hu H, Yao M, Song Y, Yang A, et al. PCAT-1 facilitates breast cancer progression by binding to RACK1 and enhancing oxygen-independent stability of HIF-1α. Mol Ther Nucleic Acids. 2021;24:310–24.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Cao J, Zhao M, Liu J, Zhang X, Pei Y, Wang J, et al. RACK1 promotes self-renewal and chemoresistance of cancer stem cells in human hepatocellular carcinoma through stabilizing Nanog. Theranostics. 2019;9:811–28.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Oien DB, Sharma S, Hattersley MM, DuPont M, Criscione SW, Prickett L, et al. BET inhibition targets ABC-DLBCL constitutive B-cell receptor signaling through PAX5. Blood Adv. 2023;7:5108–21.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Schmidt-Supprian M, Rajewsky K. Vagaries of conditional gene targeting. Nat Immunol. 2007;8:665–8.

    Article  PubMed  CAS  Google Scholar 

  30. Thévenin C, Nutt SL, Busslinger M. Early function of Pax5 (BSAP) before the pre-B-cell receptor stage of B lymphopoiesis. J Exp Med. 1998;188:735–44.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Hobeika E, Thiemann S, Storch B, Jumaa H, Nielsen PJ, Pelanda R, et al. Testing gene function early in the B-cell lineage in mb1-cre mice. Proc Natl Acad Sci USA. 2006;103:13789–94.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Nutt SL, Urbánek P, Rolink A, Busslinger M. Essential functions of Pax5 (BSAP) in pro-B-cell development: difference between fetal and adult B lymphopoiesis and reduced V-to-DJ recombination at the IgH locus. Genes Dev. 1997;11:476–91.

    Article  PubMed  CAS  Google Scholar 

  33. Arnold J, Murera D, Arbogast F, Fauny JD, Muller S, Gros F. Autophagy is dispensable for B-cell development but essential for humoral autoimmune responses. Cell Death Differ. 2016;23:853–64.

    Article  PubMed  CAS  Google Scholar 

  34. Clarke AJ, Riffelmacher T, Braas D, Cornall RJ, Simon AK. B1a B cells require autophagy for metabolic homeostasis and self-renewal. J Exp Med. 2018;215:399–413.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  35. Zhao Y, Wang Q, Qiu Q, Zhou S, Jing Z, Wang J, et al. RACK1 promotes autophagy by enhancing the Atg14L -Beclin 1-Vps34-Vps15 complex formation upon phosphorylation by AMPK. Cell Rep. 2015;13:1407–17.

    Article  PubMed  CAS  Google Scholar 

  36. Qiu G, Liu J, Cheng Q, Wang Q, Jing Z, Pei Y, et al. Impaired autophagy and defective T-cell homeostasis in mice with T-cell-specific deletion of receptor for activated C kinase 1. Front Immunol. 2017;8:575.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Rahman S, Wolberger C. Breaking the K48-chain: linking ubiquitin beyond protein degradation. Nat Struct Mol Biol. 2024;31:216–8.

    Article  PubMed  CAS  Google Scholar 

  38. Martin M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J. 2011;17:10–12.

    Article  Google Scholar 

  39. Thompson O, von Meyenn F, Hewitt Z, Alexander J, Wood A, Weightman R, et al. Low rates of mutation in clinical grade human pluripotent stem cells under different culture conditions. Nat Commun. 2020;11:1528.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Kim D, Paggi JM, Park C, Bennett C, Salzberg SL. Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype. Nat Biotechnol. 2019;37:907–15.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  41. Pertea M, Kim D, Pertea GM, Leek JT, Salzberg SL. Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Nat Protoc. 2016;11:1650–67.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15:550.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Wu T, Hu E, Xu S, Chen M, Guo P, Dai Z, et al. clusterProfiler 4.0: a universal enrichment tool for interpreting omics data. Innovation. 2021;2:100141.

    PubMed  PubMed Central  CAS  Google Scholar 

  44. Guo Y, Wang W, Wang J, Feng J, Wang Q, Jin J, et al. Zhang, Receptor for activated C kinase 1 promotes hepatocellular carcinoma growth by enhancing mitogen-activated protein kinase kinase 7 activity. Hepatology. 2013;57:140–51.

    Article  PubMed  CAS  Google Scholar 

  45. Liu G, Wang Q, Deng L, Huang X, Yang G, Cheng Q, et al. Hepatic RACK1 deficiency protects against fulminant hepatitis through myeloid-derived suppressor cells. Theranostics. 2022;12:2248–65.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  46. Cui J, Wang Q, Wang J, Lv M, Zhu N, Li Y, et al. Basal c-Jun NH2-terminal protein kinase activity is essential for survival and proliferation of T-cell acute lymphoblastic leukemia cells. Mol Cancer Ther. 2009;8:3214–22.

    Article  PubMed  CAS  Google Scholar 

  47. Ubieta K, Garcia M, Grötsch B, Uebe S, Weber GF, Stein M, et al. Fra-2 regulates B-cell development by enhancing IRF4 and Foxo1 transcription. J Exp Med. 2017;214:2059–71.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. Seo W, Ikawa T, Kawamoto H, Taniuchi I. Runx1-Cbfβ facilitates early B lymphocyte development by regulating expression of Ebf1. J Exp Med. 2012;209:1255–62.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Susan K. Pierce for providing the CD19-Cre mice, Dr. Lin Sun for providing the pCL-Eco packaging vector, and Mr. Libing Yin for FACS sorting.

Funding

This study is supported by grants from the National Natural Science Foundation of China (81930027, 92169207, and 82172298).

Author information

Author notes

  1. These authors contributed equally: Xueting Zhang, Chenke Ma.

Authors and Affiliations

  1. Beijing Institute of Basic Medical Sciences, Beijing, China

    Xueting Zhang, Chenke Ma, Yuchen Lu, Jing Wang, Hongfang Yun, Hui Jiang, Mengyao Wu, Xiaoyao Feng, Wenbin Gai, Guanglei Xu, Taoxing Shi, Qianqian Cheng & Jiyan Zhang

  2. University of South China, Hengyang Medical School, Hengyang, China

    Hui Jiang & Jiyan Zhang

  3. Anhui Medical University, Hefei, China

    Mengyao Wu & Jiyan Zhang

  4. Henan University Joint National Laboratory for Antibody Drug Engineering, Kaifeng, China

    Xiaoyao Feng, Jiannan Feng & Jiyan Zhang

  5. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China

    Hongbin Deng

  6. National Engineering Research Center for Emergence Drugs, Beijing Institute of Pharmacology and Toxicology, Beijing, China

    Jiannan Feng

  7. Institute of Life Sciences, Tsinghua University, Beijing, China

    Wanli Liu

  8. Chinese Institute for Brain Research, Beijing, China

    Jiyan Zhang

Authors
  1. Xueting Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chenke Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuchen Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hongfang Yun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hui Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mengyao Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaoyao Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Wenbin Gai
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Guanglei Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hongbin Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jiannan Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Wanli Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Taoxing Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Qianqian Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Jiyan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

JZ conceived the original ideas, designed the project, and wrote the manuscript with inputs from QC and TS. XZ, CM, JW, HY, HJ, MW, XF, GW, and GX performed the experiments. HD and WL provided key reagents. JF performed computer-guided homology modeling and molecular docking. TS, QC, and JZ supervised the investigation. XZ, CM, YL, QC, and JZ analyzed the data.

Corresponding authors

Correspondence to Taoxing Shi, Qianqian Cheng or Jiyan Zhang.

Ethics declarations

Competing interests

The authors declare that they have no competing interests. JZ is an editorial board member of Cellular & Molecular Immunology, but she has not been involved in peer review or decision-making related to the article.

Ethics approval and consent to participate

This study was approved by the Ethics Committee of the Beijing Institute of Basic Medical Sciences. The use of the clinical materials was undertaken with the understanding and signed informed consent of each donor.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, X., Ma, C., Lu, Y. et al. Rack1 regulates B-cell development and function by binding to and stabilizing the transcription factor Pax5. Cell Mol Immunol (2024). https://doi.org/10.1038/s41423-024-01213-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/s41423-024-01213-2

Keywords

Search

Advanced search

Quick links