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Identification of the Trans-Activation Domain and the Nuclear Location Signals of Human Zinc Finger Protein HZF1 (ZNF16)

Molecular Biotechnology volume 44, pages 83–89 (2010)Cite this article

Abstract

We previously characterized a C2H2-type zinc finger protein HZF1 (ZNF16) and demonstrated its important role in erythroid and megakaryocytic differentiation. This protein was located in nucleus. In this study, we first approved that HZF1 solely could activate lacZ reporter gene in yeast host Y190. This self-activation phenomenon together with structure and distribution of HZF1 suggested it as a potential transcription factor. By the auto-activation experiments and the luciferase reporter system and deletion mutation analysis, we further located the trans-activation domain at amino acid residences 49–197 within the non-zinc finger region of HZF1. An acidic residue-rich subregion (amino acids 49–105) was important for the trans-activation effect, but it could not function independently. By deletion mutation analysis, we also identified three nuclear location signals, which were located in the regions of amino acids 255–280, 328–360, and 460–490, respectively, and all of them within the zinc finger region.

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Acknowledgments

This work was supported by National Nature Science Foundation of China (30870532 and 30721063) and the Special Fund of National Laboratory of China (2060204).

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  1. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, 5 Dong Dan San Tiao, 100005, Beijing, People’s Republic of China

    Min-Jie Deng, Xiao-Bo Li, Han Peng & Jun-Wu Zhang

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  1. Min-Jie Deng
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  2. Xiao-Bo Li
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  3. Han Peng
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Corresponding author

Correspondence to Jun-Wu Zhang.

Electronic supplementary material

Supplement: Detection of nuclear location signals (NLSs) within HZF1 are shown below.

Supplement Fig. 1

Cellular localization of GFP fused with HZF1 fragment in HeLa cells transfected with partial recombination constructs. (A) DAPI staining of the HeLa cells transfected with the constructs expressing GFP fused a different HZF1 fragment. The nuclei were displayed in bright blue under fluorescent microscope at the wavelength 365 nm excitation. (B) The expression of GFP fused a HZF1 fragment in the transfected HeLa cells was observed under fluorescent microscope at the wavelength 488 nm excitation. It is noticeable that only partial cells were transformed and expressed GFP. (C) The combined result of (A) and (B) to highlight the location of GFP fused HZF1 fragment. (D) The corresponding HZF1 fragments that were fused with GFP in the recombination constructs used for the transfection experiments. (JPG 257 kb)

Supplement Fig. 2

Cellular localization of GFP fused with HZF1 fragment in HeLa cells transfected with partial recombination constructs. (A) DAPI staining of the HeLa cells transfected with the constructs expressing GFP fused a different HZF1 fragment. The nuclei were displayed in bright blue under fluorescent microscope at the wavelength 365 nm excitation. (B) The expression of GFP fused a HZF1 fragment in the transfected HeLa cells was observed under fluorescent microscope at the wavelength 488 nm excitation. It is noticeable that only partial cells were transformed and expressed GFP. (C) The combined result of (A) and (B) to highlight the location of GFP fused HZF1 fragment. (D) The corresponding HZF1 fragments that were fused with GFP in the recombination constructs used for the transfection experiments. (JPG 265 kb)

Supplement Fig. 3

Cellular localization of GFP fused with HZF1 fragment in HeLa cells transfected with partial recombination constructs. (A) DAPI staining of the HeLa cells transfected with the constructs expressing GFP fused a different HZF1 fragment. The nuclei were displayed in bright blue under fluorescent microscope at the wavelength 365 nm excitation. (B) The expression of GFP fused a HZF1 fragment in the transfected HeLa cells was observed under fluorescent microscope at the wavelength 488 nm excitation. It is noticeable that only partial cells were transformed and expressed GFP. (C) The combined result of (A) and (B) to highlight the location of GFP fused HZF1 fragment. (D) The corresponding HZF1 fragments that were fused with GFP in the recombination constructs used for the transfection experiments. (JPG 314 kb)

Supplement Fig. 4

Cellular localization of GFP fused with HZF1 fragment in HeLa cells transfected with partial recombination constructs. (A) DAPI staining of the HeLa cells transfected with the constructs expressing GFP fused a different HZF1 fragment. The nuclei were displayed in bright blue under fluorescent microscope at the wavelength 365 nm excitation. (B) The expression of GFP fused a HZF1 fragment in the transfected HeLa cells was observed under fluorescent microscope at the wavelength 488 nm excitation. It is noticeable that only partial cells were transformed and expressed GFP. (C) The combined result of (A) and (B) to highlight the location of GFP fused HZF1 fragment. (D) The corresponding HZF1 fragments that were fused with GFP in the recombination constructs used for the transfection experiments. (JPG 267 kb)

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Deng, MJ., Li, XB., Peng, H. et al. Identification of the Trans-Activation Domain and the Nuclear Location Signals of Human Zinc Finger Protein HZF1 (ZNF16). Mol Biotechnol 44, 83–89 (2010). https://doi.org/10.1007/s12033-009-9210-8

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  • DOI: https://doi.org/10.1007/s12033-009-9210-8

Keywords

  • Zinc finger protein HZF1 (ZNF16)
  • Transcriptional factor
  • Trans-activation domain
  • Nuclear location signals