Cullin-associated NEDD8-dissociated protein 1, a novel interactor of rabphilin-3A, deubiquitylates rabphilin-3A and regulates arginine vasopressin secretion in PC12 cells

Kohtaro Nakashima, Seiji Takeuchi, Shintaro Iwama, Atsushi Kiyota, Yoshinori Yasuda, Naoko Iwata, Atsushi Enomoto, Hiroshi Arima, Yoshihisa Sugimura

Research output: Contribution to journalArticle

Abstract

The molecular mechanism involved in the exocytosis of arginine vasopressin (AVP) is not fully known. Rabphilin-3A has been suggested as a novel autoantigen in infundibulo-neurohypophysitis (LINH), which leads to central diabetes insipidus through insufficient secretion of AVP. However, the role of rabphilin-3A in the pathogenesis of LINH remains unclear. Thus, the aim of the present study was to identify proteins binding rabphilin-3A in the posterior pituitary. Using glutathione S-transferase (GST)-pulldown assays and proteomic analyses, cullin-associated NEDD8-dissociated protein 1 (CAND1) was identified as a rabphilin-3A-binding protein in the posterior pituitary. Co-immunoprecipitation assays indicated that CAND1 interacted endogenously with rabphilin-3A. In addition, immunohistochemistry experiments showed that CAND1 immunoreactivity was detected mainly in the posterior pituitary, intermediate lobe, and the supraoptic nucleus in the hypothalamus, and less in the anterior lobe, partially co-localizing with rabphilin-3A. Overexpression of CAND1 resulted in deubiquitylation of rabphilin-3A in PC12 cells. Moreover, overexpression of CAND1 in PC12 cells co-transfected with AVP enhanced both basal and KCl-stimulated AVP secretion. The findings indicate that CAND1 inhibits the ubiquitylation of rabphilin-3A and positively regulates AVP secretion. These data shed light on a novel potential mechanism involving rabphilin-3A in AVP secretion, and suggest a new role of CAND1 as a regulator of hormone or neurotransmitter secretion.

Original languageEnglish
Pages (from-to)325-334
Number of pages10
JournalEndocrine Journal
Volume65
Issue number3
DOIs
Publication statusPublished - 01-01-2018

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Cullin Proteins
Arginine Vasopressin
PC12 Cells
Proteins
Neurogenic Diabetes Insipidus
rabphilin-3A
Posterior Pituitary Gland
Supraoptic Nucleus
Ubiquitination
Exocytosis
Autoantigens
Glutathione Transferase
Immunoprecipitation
Protein Binding
Proteomics
Neurotransmitter Agents
Carrier Proteins
Immunohistochemistry
Hormones

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Nakashima, Kohtaro ; Takeuchi, Seiji ; Iwama, Shintaro ; Kiyota, Atsushi ; Yasuda, Yoshinori ; Iwata, Naoko ; Enomoto, Atsushi ; Arima, Hiroshi ; Sugimura, Yoshihisa. / Cullin-associated NEDD8-dissociated protein 1, a novel interactor of rabphilin-3A, deubiquitylates rabphilin-3A and regulates arginine vasopressin secretion in PC12 cells. In: Endocrine Journal. 2018 ; Vol. 65, No. 3. pp. 325-334.
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Cullin-associated NEDD8-dissociated protein 1, a novel interactor of rabphilin-3A, deubiquitylates rabphilin-3A and regulates arginine vasopressin secretion in PC12 cells. / Nakashima, Kohtaro; Takeuchi, Seiji; Iwama, Shintaro; Kiyota, Atsushi; Yasuda, Yoshinori; Iwata, Naoko; Enomoto, Atsushi; Arima, Hiroshi; Sugimura, Yoshihisa.

In: Endocrine Journal, Vol. 65, No. 3, 01.01.2018, p. 325-334.

Research output: Contribution to journalArticle

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AU - Nakashima, Kohtaro

AU - Takeuchi, Seiji

AU - Iwama, Shintaro

AU - Kiyota, Atsushi

AU - Yasuda, Yoshinori

AU - Iwata, Naoko

AU - Enomoto, Atsushi

AU - Arima, Hiroshi

AU - Sugimura, Yoshihisa

PY - 2018/1/1

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