EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex
Abstract PAX5 and EBF1 work synergistically to regulate genes that are involved in B lymphocyte differentiation. We used the KIS-1 diffuse large B cell lymphoma cell line, which is reported to have elevated levels of PAX5 expression, to investigate the mechanism of EBF1- and PAX5-regulated gene expr...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-81000-5 |
| _version_ | 1818997780102774784 |
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| author | Charles E. Bullerwell Philippe Pierre Robichaud Pierre M. L. Deprez Andrew P. Joy Gabriel Wajnberg Darwin D’Souza Simi Chacko Sébastien Fournier Nicolas Crapoulet David A. Barnett Stephen M. Lewis Rodney J. Ouellette |
| author_facet | Charles E. Bullerwell Philippe Pierre Robichaud Pierre M. L. Deprez Andrew P. Joy Gabriel Wajnberg Darwin D’Souza Simi Chacko Sébastien Fournier Nicolas Crapoulet David A. Barnett Stephen M. Lewis Rodney J. Ouellette |
| author_sort | Charles E. Bullerwell |
| collection | DOAJ |
| description | Abstract PAX5 and EBF1 work synergistically to regulate genes that are involved in B lymphocyte differentiation. We used the KIS-1 diffuse large B cell lymphoma cell line, which is reported to have elevated levels of PAX5 expression, to investigate the mechanism of EBF1- and PAX5-regulated gene expression. We demonstrate the lack of expression of hallmark B cell genes, including CD19, CD79b, and EBF1, in the KIS-1 cell line. Upon restoration of EBF1 expression we observed activation of CD19, CD79b and other genes with critical roles in B cell differentiation. Mass spectrometry analyses of proteins co-immunoprecipitated with PAX5 in KIS-1 identified components of the MLL H3K4 methylation complex, which drives histone modifications associated with transcription activation. Immunoblotting showed a stronger association of this complex with PAX5 in the presence of EBF1. Silencing of KMT2A, the catalytic component of MLL, repressed the ability of exogenous EBF1 to activate transcription of both CD19 and CD79b in KIS-1 cells. We also find association of PAX5 with the MLL complex and decreased CD19 expression following silencing of KMT2A in other human B cell lines. These data support an important role for the MLL complex in PAX5-mediated transcription regulation. |
| first_indexed | 2024-12-20T21:51:04Z |
| format | Article |
| id | doaj.art-5b11cbd215904b91952bccb6c01327a7 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T21:51:04Z |
| publishDate | 2021-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-5b11cbd215904b91952bccb6c01327a72022-12-21T19:25:33ZengNature PortfolioScientific Reports2045-23222021-01-0111111410.1038/s41598-021-81000-5EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complexCharles E. Bullerwell0Philippe Pierre Robichaud1Pierre M. L. Deprez2Andrew P. Joy3Gabriel Wajnberg4Darwin D’Souza5Simi Chacko6Sébastien Fournier7Nicolas Crapoulet8David A. Barnett9Stephen M. Lewis10Rodney J. Ouellette11Atlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAtlantic Cancer Research Institute, Pavillon Hôtel-DieuAbstract PAX5 and EBF1 work synergistically to regulate genes that are involved in B lymphocyte differentiation. We used the KIS-1 diffuse large B cell lymphoma cell line, which is reported to have elevated levels of PAX5 expression, to investigate the mechanism of EBF1- and PAX5-regulated gene expression. We demonstrate the lack of expression of hallmark B cell genes, including CD19, CD79b, and EBF1, in the KIS-1 cell line. Upon restoration of EBF1 expression we observed activation of CD19, CD79b and other genes with critical roles in B cell differentiation. Mass spectrometry analyses of proteins co-immunoprecipitated with PAX5 in KIS-1 identified components of the MLL H3K4 methylation complex, which drives histone modifications associated with transcription activation. Immunoblotting showed a stronger association of this complex with PAX5 in the presence of EBF1. Silencing of KMT2A, the catalytic component of MLL, repressed the ability of exogenous EBF1 to activate transcription of both CD19 and CD79b in KIS-1 cells. We also find association of PAX5 with the MLL complex and decreased CD19 expression following silencing of KMT2A in other human B cell lines. These data support an important role for the MLL complex in PAX5-mediated transcription regulation.https://doi.org/10.1038/s41598-021-81000-5 |
| spellingShingle | Charles E. Bullerwell Philippe Pierre Robichaud Pierre M. L. Deprez Andrew P. Joy Gabriel Wajnberg Darwin D’Souza Simi Chacko Sébastien Fournier Nicolas Crapoulet David A. Barnett Stephen M. Lewis Rodney J. Ouellette EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex Scientific Reports |
| title | EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex |
| title_full | EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex |
| title_fullStr | EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex |
| title_full_unstemmed | EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex |
| title_short | EBF1 drives hallmark B cell gene expression by enabling the interaction of PAX5 with the MLL H3K4 methyltransferase complex |
| title_sort | ebf1 drives hallmark b cell gene expression by enabling the interaction of pax5 with the mll h3k4 methyltransferase complex |
| url | https://doi.org/10.1038/s41598-021-81000-5 |
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