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In cell lines, ATM can activate NF-\u03baB transcription factors via two independent, convergent mechanisms. One is ATM-mediated phosphorylation of nuclear NF-\u03baB essential modulator (Nemo) protein, which leads to monoubiquitylation and export of Nemo to the cytoplasm where it engages the I\u03baB kinase (IKK) complex to activate NF-\u03baB. Another is DSB-triggered migration of ATM into the cytoplasm, where it promotes monoubiquitylation of Nemo and the resulting IKK-mediated activation of NF-\u03baB. ATM has many other functions in the DSB response beyond activation of NF-\u03baB, and Nemo activates NF-\u03baB downstream of diverse stimuli, including developmental or proinflammatory stimuli such as LPSs. To elucidate the in vivo role of DSB-induced, ATM-dependent changes in expression of NF-\u03baB\u2013responsive genes, we generated mice expressing phosphomutant Nemo protein lacking consensus SQ sites for phosphorylation by ATM or related kinases. We demonstrate that these mice are viable\/healthy and fertile and exhibit overall normal B and T lymphocyte development. Moreover, treatment of their B lineage cells with LPS induces normal NF-\u03baB\u2013regulated gene expression changes. Furthermore, in marked contrast to results from a pre-B cell line, primary B lineage cells expressing phosphomutant Nemo treated with the genotoxic drug etoposide induce normal ATM- and Nemo-dependent changes in expression of NF-\u03baB\u2013regulated genes. Our data demonstrate that ATM-dependent phosphorylation of Nemo SQ motifs in vivo is dispensable for DSB-signaled changes in expression of NF-\u03baB\u2013regulated genes.<\/jats:p>","DOI":"10.4049\/jimmunol.2300139","type":"journal-article","created":{"date-parts":[[2024,7,15]],"date-time":"2024-07-15T13:10:34Z","timestamp":1721049034000},"page":"628-640","source":"Crossref","is-referenced-by-count":2,"title":["ATM-dependent Phosphorylation of Nemo SQ Motifs Is Dispensable for Nemo-mediated Gene Expression Changes in Response to DNA Double-Strand Breaks"],"prefix":"10.1093","volume":"213","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3916-6900","authenticated-orcid":false,"given":"Rebecca A","family":"Glynn","sequence":"first","affiliation":[{"name":"Cell and Molecular Biology Graduate Group, Perelman School of Medicine, University of Pennsylvania , Philadelphia, PA"},{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, Perelman School of Medicine, University of Pennsylvania , Philadelphia, PA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina E","family":"Hayer","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, Perelman School of Medicine, University of Pennsylvania , Philadelphia, PA"},{"name":"Biomedical Engineering Doctoral Degree Program, School of Biomedical Engineering, Science and Health Systems, Drexel University , Philadelphia, PA"},{"name":"Department of Biomedical and Health Informatics, Children\u2019s Hospital of Philadelphia, Perelman School of Medicine , Philadelphia, PA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1703-0786","authenticated-orcid":false,"given":"Craig H","family":"Bassing","sequence":"additional","affiliation":[{"name":"Cell and Molecular Biology Graduate Group, Perelman School of Medicine, University of Pennsylvania , Philadelphia, PA"},{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, Perelman School of Medicine, University of Pennsylvania , Philadelphia, PA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,7,15]]},"reference":[{"key":"2025012305144026600_r1","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1016\/j.cell.2017.01.002","article-title":"Endogenous DNA damage as a source of genomic instability in cancer","volume":"168","author":"Tubbs","year":"2017","journal-title":"Cell"},{"key":"2025012305144026600_r2","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.molcel.2010.09.019","article-title":"The DNA damage response: making it safe to play with knives","volume":"40","author":"Ciccia","year":"2010","journal-title":"Mol. 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