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Cloning and Functional Analysis of a Novel Human Gene ZNF415 with Multiple Isoforms

Author: ChengYingDuan
Tutor: WuXiuShan;ZhuChuanBing
School: Hunan Normal University
Course: Genetics
Keywords: ZNF415 isoform cell cycle apoptosis nucleolar protein
CLC: Q78
Type: Master's thesis
Year: 2007
Downloads: 2
Quote: 0
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Abstract


Cell cycle control is the basic activity of cell and many transcriptional factors are involved in it. Mutations and abnormal expression of the transcriptional factors will cause confusion in cell cycle. It is reported that the mutation of some transcription factors causes illness. The small count of gene number is not accord with the complex life but one gene can encode different proteins though splicing which enrich protein number greatly. The study on cell cycle factors and the isoforms is very important, we can learn basic rules of life and the causes of illness well, which is useful for disease defending and curing.With the purpose of identifying the new factors involved in cell cycle control, a new zinc finger protein was cloned from a human heart library and named ZNF415. We cloned five isoforms of this gene. These five isoforms were named ZNF415-1 to ZNF415-5. The transcriptional start sites were confirmed by RACE analysis and we found that there are some differences among the five isoforms. The five isofroms are drove by one promoter and they are occurred by alternative splicing. These isoforms display different subcellular localization, ZNF415-1 located in nucleus and ZNF415-2, -3, -4, and -5 show a nuclear and cytoplasmic expression. The five isoforms are expressed at different levels in both embryonic and adult tissues. Furthermore, the splicing variants of ZNF415display different transcriptional activities. Except for ZNF415-1, overexpression of the other ZNF415 isoforms in COS-7 cells inhibits the transcriptional activities of AP-1, p53 and p21, suggesting that the ZNF415protein may be involved in cell cycle control and/or apoptosis. Meanwhile, we use mammalian two-hybrid and co-localization to confirm the interaction with nucleolar protein HCL-G1 that was identified by yeast two-hybrid. All the isoforms may co-localized with HCL-G1, except ZNF415-1. It is reported that HCL-G1 may mediate p53 and p21 transcriptional activities and involved in rRNA splicing, suggesting that ZNF415might regulate the cell cycle/apoptosis or rRNA splicing together with HCL-G1.

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