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      Critical role of Pcid2 in B cell survival through the regulation of MAD2 expression.

      The Journal of Immunology Author Choice
      Animals, B-Lymphocytes, cytology, immunology, metabolism, Cell Cycle, genetics, Cell Cycle Proteins, biosynthesis, Cell Differentiation, Cell Survival, Gene Expression, Gene Expression Regulation, Immunoblotting, Immunohistochemistry, In Situ Hybridization, Mad2 Proteins, Mice, Mice, Knockout, RNA, Small Interfering, Reverse Transcriptase Polymerase Chain Reaction

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          Abstract

          The mitotic checkpoint is essential for maintaining genomic stability in differentiating B cells undergoing genetic alterations of the Ig gene. In this study, using real-time RT-PCR and in situ RNA hybridization, we demonstrated that MAD2 mRNA export is selectively regulated by Pcid2/Thp1. Pcid2 small interfering RNA induced a cell-cycle abnormality with increased apoptosis and polyploidy, as previously observed in MAD2-knockdown cells. Pcid2 small interfering RNA reduced MAD2 expression, but not the expression of other cell-cycle checkpoint proteins, such as MAD1 and BUBR1, or the cell-cycle-associated proteins, cyclin A, cyclin B1, and cyclin-dependent kinase 1. In mouse B lineage cells, Pcid2 transcripts appeared in a stage-dependent manner at high levels in bone marrow pre-B and immature B cells, and in spleen transitional 1 and follicular B cells, but at lower levels in pro-B, transitional 2, and marginal zone B cells, suggesting a stage-dependent requirement for MAD2 regulation. Cd19-cre-derived targeting of the Pcid2 gene induced a mature B cell deficiency in mice. These findings indicate that Pcid2 is essential for B cell survival through the regulation of MAD2 expression during B cell differentiation.

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