Analysis of PU.1/ICSBP (IRF-8) complex formation with various PU.1 mutants: molecular cloning of rat Icsbp (Irf-8) cDNA

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• 作者：Nobuhiro Nakano ; Chiharu Nishiyama ; Nobutaka Masuoka ; Makoto Nishiyama ; Hisakazu Yamane ; Ko Okumura and Hideoki Ogawa
• 关键词：cDNA ; ICSBP ; PU.1 ; Transcription factor
• 刊名：Immunogenetics
• 出版年：2005
• 出版时间：March 2005
• 年：2005
• 卷：56
• 期：12
• 页码：871-877
• 全文大小：448 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Immunology
  Allergology
  Cell Biology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1211

文摘

We isolated cDNA encoding a full-length Interferon (IFN) consensus sequence binding protein [Icsbp; also called IFN regulatory factor-8 (Irf-8)] from rat and analyzed interaction between ICSBP and PU.1, an Ets-family transcription factor regulating hematopoietic cell-specific promoters. Electrophoretic mobility shift assay indicated that PU.1 with deletion of the PEST domain could not bind to ICSBP, although loss of the PEST domain had no effect on DNA-binding ability of PU.1 itself. An amino-acid replacement of Ser147 by Ala of the PEST domain of PU.1 did not affect DNA-binding ability of PU.1 to form a binary complex, PU.1/DNA, but clearly decreased the ternary complex formation of PU.1/ICSBP/DNA. Phosphatase treatment of the ternary complex markedly decreased PU.1/ICSBP/DNA-complex formation. These results indicated that Ser147 of PU.1 is definitively required for forming a complex with ICSBP and phosphorylation of PU.1 and/or ICSBP is critical for formation of the ternary complex.