The multifunctional protein p100 is a vital transcriptional regulator that increases gene transcription by forming a physical bridge between promoter-specific transcription factors and the basal transcription machiner...The multifunctional protein p100 is a vital transcriptional regulator that increases gene transcription by forming a physical bridge between promoter-specific transcription factors and the basal transcription machinery.To identify potential signal transduction pathways in which human p100 acts as a coregulator and to find target promoter regions that may interact with p100,we performed a promoter microarray assay called chromatin immunoprecipitation-guided ligation and selection(ChIP-GLAS).From this assay,we determined that a set of promoter fragments,including several factors in the transforming growth factor beta(TGF-β)signaling pathway,exhibited interaction with p100.The ChIP-GLAS data were validated by RT-PCR assessing the mRNA expression of various factors in the TGF-b signaling pathway in cell lines.展开更多
Human p100 protein consists of four repeated domains of staphylococcal nuclease （SN）-like domain, as well as a tudor （TD） domain thereafter. We have previously shown that the SN-like domain of p100 interacted with...Human p100 protein consists of four repeated domains of staphylococcal nuclease （SN）-like domain, as well as a tudor （TD） domain thereafter. We have previously shown that the SN-like domain of p100 interacted with STAT6 and the large subunit of RNA pol Ⅱ , resulting in the enhancement of STAT6-mediated gene transcriptional activation. Here, we show that SN-like domain also interacted with CREB binding protein （CBP） and directly enhanced the acetyl transferase activity of CBP on histone. On the other hand, overexpression of CBP alone had no ability to significantly increase STAT6- dependent transcriptional activation, however, together with p100 protein, sufficiently enhanced the activation of transcription which was in line with the previous result that p100 protein bridged STAT6 with CBP.展开更多
In the present study, the interaction of human p100 protein with signal transducer and activator of transcription-6 (STAT-6) was investigated. It was proved that the staphylococcal nuclease (SN)-like and tudor (TD) do...In the present study, the interaction of human p100 protein with signal transducer and activator of transcription-6 (STAT-6) was investigated. It was proved that the staphylococcal nuclease (SN)-like and tudor (TD) domains containing in p100 protein acting as a adaptor to recruit STAT-6 to the basal transcription machinery, enhanced the STAT-6 mediated transcription activity. The interaction between STAT-6 and the p100 protein was mediated by the full-length of the SN-like domain, whereas individual fragments of SN-like domain showed no binding activity to STAT-6. In line with these results, the SN-like domain was directly engaged in the enhancement of STAT-6 mediated activation of gene transcription in vivo. Yet the TD domain had no ability to increase the transcription activation, but it was still required for the sufficient activation of transcription.展开更多
基金This work was supported by grants from the National Basic Research Program(973 Program,2009CB918903)863 Project of the Ministry of Science and Technology of China(2007AA02Z115)+3 种基金NSFC(90919032,30970562,30670441,30811130394 and 30870562)Tianjin Municipal Science and Technology Commission(08ZCGHHZ01900 and 08JCYBJC07700)Specialized Fund for the Doctoral Program of Higher Education(20091202110001)Tianjin Educational Committee Foundation(2008ZD01).
文摘The multifunctional protein p100 is a vital transcriptional regulator that increases gene transcription by forming a physical bridge between promoter-specific transcription factors and the basal transcription machinery.To identify potential signal transduction pathways in which human p100 acts as a coregulator and to find target promoter regions that may interact with p100,we performed a promoter microarray assay called chromatin immunoprecipitation-guided ligation and selection(ChIP-GLAS).From this assay,we determined that a set of promoter fragments,including several factors in the transforming growth factor beta(TGF-β)signaling pathway,exhibited interaction with p100.The ChIP-GLAS data were validated by RT-PCR assessing the mRNA expression of various factors in the TGF-b signaling pathway in cell lines.
文摘Human p100 protein consists of four repeated domains of staphylococcal nuclease （SN）-like domain, as well as a tudor （TD） domain thereafter. We have previously shown that the SN-like domain of p100 interacted with STAT6 and the large subunit of RNA pol Ⅱ , resulting in the enhancement of STAT6-mediated gene transcriptional activation. Here, we show that SN-like domain also interacted with CREB binding protein （CBP） and directly enhanced the acetyl transferase activity of CBP on histone. On the other hand, overexpression of CBP alone had no ability to significantly increase STAT6- dependent transcriptional activation, however, together with p100 protein, sufficiently enhanced the activation of transcription which was in line with the previous result that p100 protein bridged STAT6 with CBP.
文摘In the present study, the interaction of human p100 protein with signal transducer and activator of transcription-6 (STAT-6) was investigated. It was proved that the staphylococcal nuclease (SN)-like and tudor (TD) domains containing in p100 protein acting as a adaptor to recruit STAT-6 to the basal transcription machinery, enhanced the STAT-6 mediated transcription activity. The interaction between STAT-6 and the p100 protein was mediated by the full-length of the SN-like domain, whereas individual fragments of SN-like domain showed no binding activity to STAT-6. In line with these results, the SN-like domain was directly engaged in the enhancement of STAT-6 mediated activation of gene transcription in vivo. Yet the TD domain had no ability to increase the transcription activation, but it was still required for the sufficient activation of transcription.
