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Identification of Ran-binding protein M as a stanniocalcin 2 interacting protein and implications for androgen receptor activity.

Shin J, Sohn YC - BMB Rep (2014)

Bottom Line: While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1.Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner.Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

View Article: PubMed Central - PubMed

Affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung 210-702, Korea.

ABSTRACT
Stanniocalcin (STC), a glycoprotein hormone originally discovered in fish, has been implicated in calcium and phosphate homeostasis. While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1. In this study, we identified Ran-binding protein M (RanBPM) as a novel binding partner of STC2 using yeast two-hybrid screening. The interaction between STC2 and RanBPM was confirmed in mammalian cells by immunoprecipitation. STC2 enhanced the RanBPM-mediated transactivation of liganded androgen receptor (AR), but not thyroid receptor β, glucocorticoid receptor, or estrogen receptor β. We also found that AR interacted with RanBPM in both the absence and presence of testosterone (T). Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner. Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

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Related in: MedlinePlus

STC2 and RanBPM are colocalized in the cytoplasm. pcDNA3-STC2M-HIS or pcDNA3-RanBPM-AD-HA were transfected into 293T cells (A). At 48 h after transfection, the cells were fixed and incubated with HIS (for STC2M) or HA (for RanBPM-AD) antibody, followed by incubation with red-fluorescent Alexa Fluor 594 (for STC2M-HIS) (B) or green-fluorescent Alexa Fluor 488 (for RanBPM-AD-HA) (C) conjugated secondary antibody. The cells were examined by confocal laser scanning microscope. Merged and phase-contrast images (D). Number of immunoreactive or coimmunoreactive cells / number of total cells.
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Figure 0002: STC2 and RanBPM are colocalized in the cytoplasm. pcDNA3-STC2M-HIS or pcDNA3-RanBPM-AD-HA were transfected into 293T cells (A). At 48 h after transfection, the cells were fixed and incubated with HIS (for STC2M) or HA (for RanBPM-AD) antibody, followed by incubation with red-fluorescent Alexa Fluor 594 (for STC2M-HIS) (B) or green-fluorescent Alexa Fluor 488 (for RanBPM-AD-HA) (C) conjugated secondary antibody. The cells were examined by confocal laser scanning microscope. Merged and phase-contrast images (D). Number of immunoreactive or coimmunoreactive cells / number of total cells.

Mentions: To examine the subcellular localizations of STC2 and RanBPM, 293T cells were cotransfected with the STC2M-HIS and RanBPM-AD-HA expression vectors followed by indirect immunofluorescence detection (Fig. 2A). The red fluorescence of STC2M-HIS showed strong immunoreactivity in the cytoplasm (Fig. 2B). Immunoreactivity for RanBPM-AD-HA was observed in both the nucleus and cytoplasm (Fig. 2C). Merged images showed that STC2 and RanBPM were co-localized in the cytoplasm (Fig. 2D).


Identification of Ran-binding protein M as a stanniocalcin 2 interacting protein and implications for androgen receptor activity.

Shin J, Sohn YC - BMB Rep (2014)

STC2 and RanBPM are colocalized in the cytoplasm. pcDNA3-STC2M-HIS or pcDNA3-RanBPM-AD-HA were transfected into 293T cells (A). At 48 h after transfection, the cells were fixed and incubated with HIS (for STC2M) or HA (for RanBPM-AD) antibody, followed by incubation with red-fluorescent Alexa Fluor 594 (for STC2M-HIS) (B) or green-fluorescent Alexa Fluor 488 (for RanBPM-AD-HA) (C) conjugated secondary antibody. The cells were examined by confocal laser scanning microscope. Merged and phase-contrast images (D). Number of immunoreactive or coimmunoreactive cells / number of total cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4281344&req=5

Figure 0002: STC2 and RanBPM are colocalized in the cytoplasm. pcDNA3-STC2M-HIS or pcDNA3-RanBPM-AD-HA were transfected into 293T cells (A). At 48 h after transfection, the cells were fixed and incubated with HIS (for STC2M) or HA (for RanBPM-AD) antibody, followed by incubation with red-fluorescent Alexa Fluor 594 (for STC2M-HIS) (B) or green-fluorescent Alexa Fluor 488 (for RanBPM-AD-HA) (C) conjugated secondary antibody. The cells were examined by confocal laser scanning microscope. Merged and phase-contrast images (D). Number of immunoreactive or coimmunoreactive cells / number of total cells.
Mentions: To examine the subcellular localizations of STC2 and RanBPM, 293T cells were cotransfected with the STC2M-HIS and RanBPM-AD-HA expression vectors followed by indirect immunofluorescence detection (Fig. 2A). The red fluorescence of STC2M-HIS showed strong immunoreactivity in the cytoplasm (Fig. 2B). Immunoreactivity for RanBPM-AD-HA was observed in both the nucleus and cytoplasm (Fig. 2C). Merged images showed that STC2 and RanBPM were co-localized in the cytoplasm (Fig. 2D).

Bottom Line: While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1.Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner.Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

View Article: PubMed Central - PubMed

Affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung 210-702, Korea.

ABSTRACT
Stanniocalcin (STC), a glycoprotein hormone originally discovered in fish, has been implicated in calcium and phosphate homeostasis. While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1. In this study, we identified Ran-binding protein M (RanBPM) as a novel binding partner of STC2 using yeast two-hybrid screening. The interaction between STC2 and RanBPM was confirmed in mammalian cells by immunoprecipitation. STC2 enhanced the RanBPM-mediated transactivation of liganded androgen receptor (AR), but not thyroid receptor β, glucocorticoid receptor, or estrogen receptor β. We also found that AR interacted with RanBPM in both the absence and presence of testosterone (T). Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner. Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

Show MeSH
Related in: MedlinePlus