Role for Btg1 and Btg2 in growth arrest of WEHI-231 cells through arginine methylation following membrane immunoglobulin engagement.
Engagement of membrane Ig (mIg) on WEHI-231 murine B lymphoma cells, a cell line model representative of primary immature B cells, results in growth arrest and subsequent apoptosis. Of the several dozen genes upregulated greater than two-fold by anti-IgM treatment through DNA microarray analysis, we focused on B cell translocation gene 1 (Btg1) and Btg2, member of Btg/Tob family of proteins. WEHI-231 cells were infected with the Btg1/EGFP or Btg2/EGFP retroviral vectors, and those expressing either Btg1 or Btg2 accumulated in G1 phase at significantly higher proportions than that seen for cells expressing control vector. Btg1 or Btg2 bound to protein arginine methyltransferase (PRMT) 1 via the box C region, an interaction required for anti-IgM-induced growth inhibition. The arginine methyltransferase inhibitor AdOx partially abrogated growth inhibition induced by Btg1, Btg2, or anti-IgM. The Btg1- or Btg2-induced growth inhibition was also abrogated in PRMT1-deficient cells via introduction of small interference RNA. In addition, we observed anti-IgM-induced arginine methylation of two proteins, a 28-kDa and a 36-kDa protein. Methylation, detected by a monoclonal antibody specific for asymmetric, but not symmetric methyl residues, was observed as early as 1 h-2 h after stimulation and was sustained for up to 24 h. The anti-IgM-induced p36 arginine methylation was abrogated in the PRMT1-deficient cells, suggesting that PRMT1 induces p36 methylation. Together, these results suggest that anti-IgM-induced growth inhibition is mediated via upregulation of Btg1 and Btg2, resulting in the activation of arginine methyltransferase activity and culminating in growth inhibition of WEHI-231 cells.
Hata
Kikumi
K
Department of Immunology and Intractable Immunology Research Center, Tokyo Medical University, 6-1-1 Shinjuku, Tokyo, Japan.
Nishijima
Kana
K
Mizuguchi
Junichiro
J
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D001120:Arginine / Q000378:metabolism*; D001402:B-Lymphocytes / Q000276:immunology*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000502:physiology; D002462:Cell Membrane / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D004252:DNA Mutational Analysis; D016193:G1 Phase / Q000187:drug effects; D016147:Genes, Tumor Suppressor; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D007075:Immunoglobulin M / Q000032:analysis / Q000187:drug effects*; D047908:Intracellular Signaling Peptides and Proteins; D008745:Methylation / Q000187:drug effects; D008780:Methyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics; D051379:Mice; D009025:Morpholines / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011484:Protein-Arginine N-Methyltransferases; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics; D017384:Sequence Deletion; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins; D015854:Up-Regulation
D000888:Antibodies, Anti-Idiotypic; C489552:Btg1 protein, mouse; C489435:Btg2 protein, mouse; D018797:Cell Cycle Proteins; D002867:Chromones; D017874:Immediate-Early Proteins; D007075:Immunoglobulin M; D047908:Intracellular Signaling Peptides and Proteins; D009025:Morpholines; D009363:Neoplasm Proteins; D000081082:Phosphoinositide-3 Kinase Inhibitors; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C485732:Tob2 protein, mouse; D025521:Tumor Suppressor Proteins; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C017199:2',3'-dialdehyde ATP; D000255:Adenosine Triphosphate; D001120:Arginine; D008780:Methyltransferases; C077013:PRMT2 protein, human; D011484:Protein-Arginine N-Methyltransferases