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Fank1 interacts with Jab1 and regulates cell apoptosis via the AP-1 pathway

Cellular and Molecular Life Sciences volume 68, pages 2129–2139 (2011)Cite this article

Abstract

Regulation of apoptosis at various stages of differentiation plays an important role in spermatogenesis. Therefore, the identification and characterisation of highly expressed genes in the testis that are involved in apoptosis is of great value to delineate the mechanism of spermatogenesis. Here, we reported that Fank1, a novel gene highly expressed in testis, functioned as an anti-apoptotic protein that activated the activator protein 1 (AP-1) pathway. We found that Jab1 (Jun activation domain-binding protein 1), a co-activator of AP-1, specifically interacted with Fank1. Reporter analyses showed that Fank1 activated AP-1 pathway in a Jab1-dependent manner. Fank1 overexpression also increased the expression and activation of endogenous c-Jun. Further study showed that Fank1 inhibited cell apoptosis by upregulating and activating endogenous c-Jun and its downstream target, Bcl-3. This process was shown to be Jab1 dependent. Taken together, our results indicated that by interacting with Jab1, Fank1 could suppress cell apoptosis by activating the AP-1-induced anti-apoptotic pathway.

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Acknowledgments

We thank Dr. Dong-Yan Jin for constructive suggestions and critical reading of manuscript. This work was supported by grants from the National Program for Important Research Plans (2006CB944002) and the National Program for Key Basic Research Projects (2006CB504001) from the Ministry of Science and Technology of China, a grant for Creative Research Group (No. 30721063) from the National Natural Science Foundation of China, the National Laboratory Special Fund (2060204), and the Institute Fund for Young Scientist (2009PY10).

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Authors and Affiliations

  1. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Tsinghua University, 5 Dong Dan San Tiao, Beijing, 100005, China

    Hailong Wang, Wei Song, Tinghui Hu, Ning Zhang, Shiying Miao & Linfang Wang

  2. Academy of Basic Medicine, Shanxi Medical University, 56 Xin Jian Nan Lu, Taiyuan, 030001, China

    Hailong Wang

  3. National Research Institute for Family Planning Beijing, WHO Collaboration Center of Human Reproduction, 12 Da Hui Si, Hai Dian, Beijing, 100081, China

    Shudong Zong

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  1. Hailong Wang
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  2. Wei Song
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  3. Tinghui Hu
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Correspondence to Wei Song or Linfang Wang.

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Supplementary Fig. 1

PI incorporation FACS analysis of Fank1 inhibition of CPT induced apoptosis. Stable HeLa/Flag-Fank1 and HeLa/Con cell lines were exposed to 10 µM CPT. At indicated times, cells were treated as described in ‘Materials and Methods’ section and analysed by FACS. The percentage of apoptotic cells (% of total cells) was determined by program EXPO32-ADC, and the results were shown as a bar graph (*p<0.05). The data were representative of three different experiments and error bars represented the standard deviations of triplicate samples. (TIFF 19882 kb)

Supplementary Fig. 2

Fank1 inhibited CPT induced apoptosis by upregulating c-Jun and Bcl-3 expression. Stable HeLa/Flag-Fank1 and HeLa/Con cell lines were exposed to 10 µM CPT. The cell lysates were analysed by immunoblotting with the indicated antibodies. GAPDH was used as a loading control. (TIFF 24484 kb)

Supplementary Fig. 3

Annexin V/PI FACS assay to analyse apoptosis in stable cells transfected with the indicated siRNA (Jab1 siRNA2 or control siRNA). Stable cells were treated as described in ‘Materials and Methods’. After staining with FITC-Annexin V and PI, cells were analysed by the Accuri C6 flow cytometer system. Inset numbers represent the percentage of each population in the quadrants. Early apoptotic cells (Annexin V+/PI-, Q1-LR), late apoptotic cells (Annexin V+/PI+, Q1-UR) and dead cells (Annexin V-/PI+, Q1-UL). Data were from one of three independent experiments which yielded similar results. (TIFF 25386 kb)

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Wang, H., Song, W., Hu, T. et al. Fank1 interacts with Jab1 and regulates cell apoptosis via the AP-1 pathway. Cell. Mol. Life Sci. 68, 2129–2139 (2011). https://doi.org/10.1007/s00018-010-0559-4

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  • DOI: https://doi.org/10.1007/s00018-010-0559-4

Keywords

  • Fank1
  • Jab1
  • AP-1
  • Bcl-3
  • Apoptosis