腦指蛋白 (Znf179)結合早幼粒細胞白血病鋅指蛋白 (PLZF)，並在蛋白質層面調控其基因表現

謝亞庭

doi:10.6844/NCKU.2010.01349

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腦指蛋白 (Znf179)結合早幼粒細胞白血病鋅指蛋白 (PLZF)，並在蛋白質層面調控其基因表現

Znf179 interacts with PLZF and regulates PLZF gene expression at protein level

謝亞庭 , Masters　　Advisor：李宜釗　　

繁體中文 DOI： 10.6844/NCKU.2010.01349

腦指蛋白；早幼粒細胞白血病鋅指蛋白；酵母雙雜合系統； Znf179 ； Promyelocytic Leukemia Zinc-Finger (PLZF) ； Yeast two-hybrid

Abstract │ Reference (72) │ Altmetrics

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Znf179 is a member of the RING finger protein family. During embryogenesis, Znf179 is restrictedly expressed in brain region, suggesting a potential role in neuronal development. Previous studies in our laboratory have demonstrated that the expression of Znf179 is remarkably increased in the retinoic acid-induced neuronal differentiated P19 cells and Znf179 knock-down significantly attenuates neuronal differentiation. To investigate the molecular mechanisms of Znf179 function, we performed yeast two-hybrid screen to identify the Znf179 interacting proteins. In the experiments, promyelocytic leukemia zinc-finger (PLZF) was identified as a novel Znf179 interacting protein. The results of immunoprecipitation assay also confirmed the interaction between PLZF and Znf179 in vivio. Furthermore, we have demonstrated that PLZF interacts with Znf179 via its two zinc-finger domains, which is located between amino acids 398 and 455. Using immunofluorescence analysis, we observed that the cellular localization of Znf179 was changed by co-expressing PLZF. Taken together, these results indicate that the two proteins dose indeed interact with each other in vivo. PLZF has been found to regulate transcription through recruitment of nuclear receptor corepressors (N-CoR or SMRT) / histone deacetylase (HDAC) complexes via its POZ domain. To investigate whethere Znf179 affects transcriptional regulation ability of PLZF, COS-1 cells were co-transfected with HA-PLZF and EGFP-Znf179 or control plasmid. Unexpectedly, we found that exogenously expressing Znf179 caused a 5-fold increase in HA-PLZF protein abundance but not at Flag-PLZF RNA level. Further studies will be performed to examine the molecular mechanisms of Znf179 functions on PLZF protein regulation.

Reference ( 72 ) 〈TOP〉

1. Inoue S, Orimo A, Saito T, Ikeda K, Sakata K, Hosoi T, Orimo H, Ouchi Y, Muramatsu M. A novel RING finger protein, BFP, predominantly expressed in the brain. Biochem Biophys Res Commun 1997 Nov 7; 240(1):8-14.
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2. Saurin AJ, Borden KL, Boddy MN, Freemont PS. Does this have a familiar RING? Trends Biochem Sci 1996 Jun; 21(6):208-14.
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3. Borden, K.L. & Freemont, P.S. The RING finger domain: a recent example of a sequence-structure family. Curr Opin Struct Biol 1996 Jun; 6(3):395-401.
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4. Deshaies, R. J., Joazeiro, C. A. RING domain E3 ubiquitin ligases. Annu Rev Biochem 2009; 78:399-434.
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5. Orimo A, Inoue S, Ikeda K, Sato M, Kato A, Tominaga N, Suzuki M, Noda T, Watanabe M, Muramatsu M. Molecular cloning, localization, and developmental expression of mouse brain finger protein (Bfp)/ZNF179: distribution of bfp mRNA partially coincides with the affected areas of Smith-Magenis syndrome. Genomics 1998 Nov 15; 54(1):59-69.
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