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ATAT1 is essential for regulation of homeostasis-retaining cellular responses in corticotrophs along hypothalamic-pituitary-adrenal axis

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Abstract

The production and secretion of adrenocorticotropin, a proopiomelanocortin (POMC)-derived hormone, by corticotrophs in the anterior pituitary, is regulated by corticotrophin-releasing hormone (CRH) and glucocorticoids. We have previously demonstrated that adrenalectomy induces α-tubulin N-acetyltransferase 1 (ATAT1) expression and α-tubulin acetylation in corticotrophs. However, the regulatory mechanism of ATAT1 expression and the function of acetylated microtubules in corticotrophs are unclear. Here, we analyze the effect of CRH or dexamethasone on Atat1 expression in the mouse corticotroph AtT20 cell line. The expression of Atat1 was increased by CRH and decreased by dexamethasone in AtT20 cells. We examined the effect of Atat1 knockdown on the expression of POMC-associated genes and the dexamethasone-induced nuclear translocation of glucocorticoid receptor (GR) by real-time polymerase chain reaction and Western blot analysis, respectively. Atat1 knockdown resulted in a significant increase in the expression of ACTH-producing genes and decreased the dexamethasone-induced nuclear translocation of GR accompanied with a reduction in α-tubulin acetylation. Atat1 overexpression resulted in a significant increase in α-tubulin acetylation and the dexamethasone-induced nuclear translocation of GR. These results suggest that the acetylated microtubules function as the rail-line for the transportation of GR into the nucleus. We conclude that ATAT1 finely tunes the cellular responses of corticotrophs to hormonal stimulation through an intracellular feedback circuit.

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Acknowledgments

This work was supported in part by a Grant-in-Aid for Young Scientists (B) (16 K18982) to T. N. from the Ministry of Education, Culture, Sports, Science and Technology of Japan, by a grant from the Yuasa Memorial Foundation and by a research grant from Teikyo University School of Medicine. We thank Dr. Masayoshi Iizuka (Teikyo University School of Medicine, Tokyo, Japan) for excellent technical advice and Editage (www.editage.jp) for English language editing.

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Teikyo University School of Medicine, 2-11-1 Kaga Itabashi-Ku, Tokyo, 173-8605, Japan

    Takashi Nakakura, Anshin Asano-Hoshino, Yoko Nekooki-Machida, Hideyuki Tanaka, Kenjiro Arisawa, Yoshimi Nishijima, Yoshiko Kiuchi & Haruo Hagiwara

  2. Department of Biology, Sapporo Medical University, Sapporo, Japan

    Takeshi Suzuki

  3. Biosignal Research Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Japan

    Seiji Torii

  4. Department of Biochemistry, Teikyo University School of Medicine, Tokyo, Japan

    Takao Susa & Tomoki Okazaki

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  1. Takashi Nakakura
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Correspondence to Takashi Nakakura.

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Supplemental Fig. 1

Acetylation levels of HSP90 (a) and cytoplasmic dynein (b) in siAtat1- and NT-transfected AtT20 cells. GR has been reported to form a protein complex with HSP90 (a chaperone protein) and cytoplasmic dynein (Grad and Picard 2007). Although HSP90 is deacetylated by HDAC6 (Li et al. 2013), whether ATAT1 is involved in HSP90 acetylation remains unclear. We examined the acetylation level of HSP90 in siAtat1- and NT-transfected AtT20 cells by Western blot analysis. a Acetylated and total HSPs were detected with rabbit anti-acetylated HSP90 (1:2500; 600–401-981, Rockland, Pa., USA) and mouse anti-HSP90 (1:2500; ADI-SPA-830, Enzo life science, N.Y., USA), respectively. Atat1 knockdown showed no effect on both acetylation and expression level of HSP90. b As cytoplasmic dynein functions as a motor protein for the transport of the GC/GR complex from cytoplasm to nucleus along MTs, we compared the cytoplasmic dynein levels in siAtat1- and NT-transfected cells by Western blot analysis with mouse anti-dynein (1:5000; 14–9772-82; eBioscience, Calif., USA). The level of cytoplasmic dynein was the same in both treatment groups. (GIF 37 kb)

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Nakakura, T., Suzuki, T., Torii, S. et al. ATAT1 is essential for regulation of homeostasis-retaining cellular responses in corticotrophs along hypothalamic-pituitary-adrenal axis. Cell Tissue Res 370, 169–178 (2017). https://doi.org/10.1007/s00441-017-2654-4

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