Induction of light chain replacement in human plasma cells by caffeine is independent from both the upregulation of RAG protein expression and germ line transcription.
When some human plasma cell lines are cultured with concanavalin A, the original light chain is replaced with another light chain which results from secondary VJ recombination (light chain shifting). We examined various intracellular factors involved in the induction of light chain shifting. Light chain shifting can be induced upon treatment with agents with phosphatase inhibitory activity such as caffeine and okadaic acid. Although the plasma cells used express both RAG-1 and RAG-2, the expression level of these proteins was not affected by caffeine or okadaic acid. Transcription of the germ line locus, which correlates to the locus activation for rearrangement, is also not influenced by phosphatase inhibition. However, the amount of signal broken-ended DNA intermediates generated during V(D)J rearrangement was shown to increase upon caffeine or okadaic acid treatment. The inhibitory activity of caffeine on phosphatase was the same as okadaic acid. However, caffeine exhibited much higher activity for VJ coding joint formation than okadaic acid. Therefore, although phosphatase inhibition might act, in part, on a mechanism by which V(D)J recombinase activity is regulated within the human plasma cells, other factor(s) are probably also involved in the process.
Tachibana
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H
Division of Bioresources, Graduate School, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka, 812-8581, Japan. tatibana@agr.kyushu-u.ac.jp
Haruta
H
H
Ueda
K
K
Chiwata
T
T
Yamada
K
K
D001483:Base Sequence; D015153:Blotting, Western; D002110:Caffeine / Q000494:pharmacology*; D002460:Cell Line; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016654:Genes, RAG-1 / Q000235:genetics*; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D019319:Okadaic Acid / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors; D010950:Plasma Cells / Q000378:metabolism*; D011995:Recombination, Genetic / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation; D045567:VDJ Recombinases
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D018398:Homeodomain Proteins; D007147:Immunoglobulin Light Chains; D009687:Nuclear Proteins; D010726:Phosphodiesterase Inhibitors; C499311:RAG2 protein, human; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D019319:Okadaic Acid; D002110:Caffeine; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases; D010744:Phosphoric Monoester Hydrolases