一种转录谱分析方法揭示锌指转录因子ZNF191是一种多效性因子。
A transcript profiling approach reveals the zinc finger transcription factor ZNF191 is a pleiotropic factor.
作者信息
Li Jianzhong, Chen Xia, Gong Xuelian, Liu Ying, Feng Hao, Qiu Lei, Hu Zhenlin, Zhang Junping
机构信息
Department of Biochemical Pharmacy, Second Military Medical University, Shanghai, PR China.
出版信息
BMC Genomics. 2009 May 22;10:241. doi: 10.1186/1471-2164-10-241.
BACKGROUND
The human zinc finger protein 191 (ZNF191) is a member of the SCAN domain family of Krüppel-like zinc finger transcription factors. ZNF191 shows 94% identity to its mouse homologue zinc finger protein 191(Zfp191), which is the most highly conserved among the human-mouse SCAN family member orthologues pairs. Zfp191 is widely expressed during early embryogenesis and in adult organs. Moreover, Zfp191-/- embryos have been shown to be severely retarded in development and die approximately at embryonic day E7.5. ZNF191 can specifically interact with the widespread TCAT motif which constitutes the HUMTH01 microsatellite in the tyrosine hydroxylase (TH) gene. Allelic variations of HUMTH01 have been stated to have a quantitative silencing effect on TH gene expression and to correlate with quantitative and qualitative changes in the binding by ZNF191. In addition, ZNF191 displays a suppressive effect on the transcription; however, little downstream targets have identified.
RESULTS
We searched for ZNF191 target genes by using a transient overexpression and knockdown strategy in the human embryo kidney (HEK293) cells. Microarray analyses identified 6094 genes modulated by overexpression of ZNF191 and 3332 genes regulated by knockdown of ZNF191, using a threshold of 1.2-fold. Several interested candidate genes, validated by real time RT-PCR, were correlated well with the array data. Interestingly, 1456 genes were identified in both transient overexpression and transient knockdown strategies. The GenMAPP and MappFinder software packages were further used for pathway analysis of these significantly altered genes. Several gene pathways were found to be involved in processes of the regulation of kinase activity, transcription, angiogenesis, brain development and response to DNA damage.
CONCLUSION
Our analysis reveals for the first time that ZNF191 is a pleiotropic factor that has a role in hematopoiesis, brain development and cancers.
背景
人类锌指蛋白191（ZNF191）是Krüppel样锌指转录因子的SCAN结构域家族成员。ZNF191与其小鼠同源物锌指蛋白191（Zfp191）具有94%的同一性，这在人-小鼠SCAN家族成员直系同源物对中是保守性最高的。Zfp191在胚胎发育早期和成年器官中广泛表达。此外，已证明Zfp191基因敲除的胚胎发育严重迟缓，大约在胚胎第7.5天死亡。ZNF191可特异性地与构成酪氨酸羟化酶（TH）基因中HUMTH01微卫星的广泛存在的TCAT基序相互作用。据报道，HUMTH01的等位基因变异对TH基因表达具有定量沉默作用，并与ZNF191结合的定量和定性变化相关。此外，ZNF191对转录具有抑制作用；然而，已鉴定的下游靶点很少。
结果
我们在人胚肾（HEK293）细胞中采用瞬时过表达和敲低策略寻找ZNF191的靶基因。微阵列分析确定，使用1.2倍的阈值，有6094个基因受ZNF191过表达调节，3332个基因受ZNF191敲低调节。通过实时逆转录-聚合酶链反应验证的几个感兴趣的候选基因与阵列数据相关性良好。有趣的是，在瞬时过表达和瞬时敲低策略中均鉴定出1456个基因。进一步使用GenMAPP和MappFinder软件包对这些显著改变的基因进行通路分析。发现几个基因通路参与激酶活性调节、转录、血管生成、脑发育和对DNA损伤的反应过程。
结论
我们的分析首次揭示ZNF191是一种多效性因子，在造血、脑发育和癌症中发挥作用。
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