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首页> 中文期刊> 《中国病理生理杂志》 >转录因子GATA-1与人巨细胞病毒UL111A基因5'上游序列结合的实验研究

转录因子GATA-1与人巨细胞病毒UL111A基因5'上游序列结合的实验研究

         
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摘要

ATM; To investigate the interactions between transcription factor GATA -1 and 5' upstream DNA sequence of human cytomegalovirus ( HCMV) UL111A gene using established HCMV latent and reactivated infection mod -els. METHODS: The expression of transcription factor GATA - 1 in the process of HCMV latent and reactivated infections was determined by Western blotting. Sequence analysis revealed the presence of 11 consensus GATA -1 binding sites on the 5 'upstream DNA sequence of UL111A gene using MATCH tool software . Moreover, 9 pairs of primers were designed for the core DNA sequence of the 11 binding sites. The bindings of GATA - 1 with 5' upstream DNA sequence of UL111A gene in HCMV latent and reactivated infections were assessed by real - time fluorescence quantitative PCR and chromatin immu -noprecipitation ( ChIP). RESULTS: UL111A gene did not express cmvIL - 10 in latent infection but did in reactivated in -fection. The relative expression of transcription factor GATA - 1 was more in HCMV latent infection than that in reactivated infection. Five pairs of primers were used to amplify the target band when using the GATA-1 antibody — specific bindingrnDNA fragments as templates. The binding rates of GATA -1 of 5 primers in HCMV latent infection group were - 4. 00 ± 0.26, -4.50±0.33, -4.41 ±0.21, -3.61 ±0.20 and -3.47 ±0.11 , while that in reactivated infection group were -1.13±0.07, -0.63±0.05, -1.10±0.07, - 0. 24 ± 0. 03 and -0.14 ±0.01 (P<0.05). CONCLUSION: Transcription factor GATA -1 mediates the UL111A transcription which is involved in HCMV latent and reactivated infec -tions by binding the HCMV UL111A gene 5' upstream sequence 107 ( -4772) site, 609 ( -4270) site, 849 ( -4030) site, 1119 ( -3760) site and 2047 ( -2832) site during reactivated infection.%目的:利用人巨细胞病毒(human cytomegalovirus,HCMV)潜伏及再激活感染THP-1细胞模型,研究细胞核内转录因子GATA-1与HCMV UL111A基因5上游DNA序列的相互作用.方法:采用免疫印迹(Western blotting)技术定量分析HCMV潜伏及再激活感染时细胞核内转录因子GATA-1的表达;通过MATCH工具软件分析UL111A基因5上游DNA序列,发现存在着11个GATA-1的结合位点,针对这11个GATA-1结合位点DNA序列设计9对引物,采用染色质免疫沉淀(chromatin immunoprecipitation,ChIP)及实时荧光定量PCR技术分析转录因子GATA-1与UL111A基因5上游DNA序列的结合情况.结果:HCMV潜伏感染时UL111A基因未表达cmvIL-10,激活感染时表达cmvIL-10,潜伏感染时转录因子GATA-1的相对表达量高于激活感染;以GATA-1抗体特异性结合的DNA片段为模板,9对引物中共有5对扩增出目的条带;HCMV潜伏感染组和激活感染组5个位点转录因子GATA-1结合率分别为:-4.00±0.26、-4.50±0.33、-4.41±0.21、-3.61±0.20、-3.47±0.11和-1.13±0.07、-0.63±0.05、-1.10±0.07、-0.24±0.03和-0.14±0.01,差异均有统计学意义(P<0.05).结论:HCMV感染时转录因子GATA-1可能通过与UL111A基因 5'上游序列1119(-3760)位点、107(-4772)位点、609(-4270)位点、849(-4030)位点和2047(-2832)位点结合,调节UL111A基因的转录,参与HCMV潜伏与再激活感染的过程.

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  • 来源
    《中国病理生理杂志》 |2012年第12期|2244-2249|共6页
  • 作者

    田可港; 浮苗; 高艳; 彭颖; 李祥; 郑晓群; 徐志伟; 宋建华; 吕建新;

  • 作者单位

    宁波市鄞州人民医院;

    温州医学院附属第二医院实验诊断中心,浙江,温州,325027;

    温州医学院附属第二医院实验诊断中心,浙江,温州,325027;

    温州医学院浙江省医学遗传学重点实验室,浙江,温州,325035;

    温州医学院浙江省医学遗传学重点实验室,浙江,温州,325035;

    温州医学院附属第二医院实验诊断中心,浙江,温州,325027;

    温州医学院附属第二医院实验诊断中心,浙江,温州,325027;

    俄克拉荷马医学研究中心,美国,俄克拉荷马,73104;

    温州医学院浙江省医学遗传学重点实验室,浙江,温州,325035;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 其他病毒;
  • 关键词

    转录因子GATA-1; HCMV潜伏感染; UL111A;

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