1,25-Dihydroxyvitamin D3 translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells

Maura Simboli-Campbell; Anne Marie Gagnon; Douglas J. Franks; Jo Ellen Welsh

doi:10.1016/s0021-9258(17)41856-4

1,25-Dihydroxyvitamin D₃ translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells

Maura Simboli-Campbell
, Anne Marie Gagnon
, Douglas J. Franks
, Jo Ellen Welsh

Environmental Health Sciences

University at Albany

University of Ottawa

Research output: Contribution to journal › Article › peer-review

64 Scopus citations

Abstract

1,25-Dihydroxycholecalciferol (1,25-(OH)₂-D₃) increases membrane- associated protein kinase C (PKC) activity and immunoreactivity in renal epithelial (Madin Darby bovine kidney, MDBK) cells (Simboli-Campbell, M., Franks, D. J., and Welsh, J. E. (1992) Cell Signalling 4, 99-109). We have now characterized the effects of 1,25-(OH)₂-D₃ on the subcellular localization of three individual isozymes by immunofluorescence and immunoblotting. Although the total amount of PKCα, PKCβ, and PKCζ are unaffected by 1,25-(OH)₂-D₃, this steroid hormone induces subcellular redistribution of both PKCα and PKCβ. Treatment with 1,25-(OH)₂-D₃ (100 nM, 24 h) enhances plasma membrane association of PKCα and induces translocation of PKCβ to the nuclear membrane. The effects of 1,25-(OH)₂- D₃ appear to be limited to the calcium-dependent PKC isozymes, since 1,25- (OH)₂-D₃ has no effect on the calcium independent isozyme, PKCζ. In contrast to rapid transient PKC translocation seen in response to agents which interact with membrane receptors to induce phospholipid hydrolysis, modulation of PKCα and PKCβ is observed after 24 h treatment with 1,25- (OH)₂-D₃. In MDBK cells, the phorbol ester 12-O-tetradecanoylphorbol-13- acetate (TPA) (100 nM, 24 h) down-regulates PKCα and, to a lesser extent, PKCζ, without altering their subcellular distribution. TPA also induces translocation of PKCβ to the nuclear membrane. MDBK cells treated with 1,25- (OH)₂-D₃, but not TPA, exhibit enhanced phosphorylation of endogenous nuclear proteins. In addition to the distinct effects of 1,25-(OH)₂-D₃ and TPA on PKC isozyme patterns, 1,25-(OH)₂-D₃ up-regulates both the vitamin D receptor and calbindin D-28K, whereas TPA down-regulates the expression of both proteins. These data support the involvement of PKC in the mechanism of action of 1,25-(OH)₂-D₃ and specifically implicate PKCβ in 1,25-(OH)₂- D₃-mediated nuclear events.

Original language	English
Pages (from-to)	3257-3264
Number of pages	8
Journal	Journal of Biological Chemistry
Volume	269
Issue number	5
DOIs	https://doi.org/10.1016/s0021-9258(17)41856-4
State	Published - Feb 4 1994

Access to Document

10.1016/s0021-9258(17)41856-4

Cite this

@article{5f7e83b9b29f48cabf2381886e2b5c5a,

title = "1,25-Dihydroxyvitamin D3 translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells",

abstract = "1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) increases membrane- associated protein kinase C (PKC) activity and immunoreactivity in renal epithelial (Madin Darby bovine kidney, MDBK) cells (Simboli-Campbell, M., Franks, D. J., and Welsh, J. E. (1992) Cell Signalling 4, 99-109). We have now characterized the effects of 1,25-(OH)2-D3 on the subcellular localization of three individual isozymes by immunofluorescence and immunoblotting. Although the total amount of PKCα, PKCβ, and PKCζ are unaffected by 1,25-(OH)2-D3, this steroid hormone induces subcellular redistribution of both PKCα and PKCβ. Treatment with 1,25-(OH)2-D3 (100 nM, 24 h) enhances plasma membrane association of PKCα and induces translocation of PKCβ to the nuclear membrane. The effects of 1,25-(OH)2- D3 appear to be limited to the calcium-dependent PKC isozymes, since 1,25- (OH)2-D3 has no effect on the calcium independent isozyme, PKCζ. In contrast to rapid transient PKC translocation seen in response to agents which interact with membrane receptors to induce phospholipid hydrolysis, modulation of PKCα and PKCβ is observed after 24 h treatment with 1,25- (OH)2-D3. In MDBK cells, the phorbol ester 12-O-tetradecanoylphorbol-13- acetate (TPA) (100 nM, 24 h) down-regulates PKCα and, to a lesser extent, PKCζ, without altering their subcellular distribution. TPA also induces translocation of PKCβ to the nuclear membrane. MDBK cells treated with 1,25- (OH)2-D3, but not TPA, exhibit enhanced phosphorylation of endogenous nuclear proteins. In addition to the distinct effects of 1,25-(OH)2-D3 and TPA on PKC isozyme patterns, 1,25-(OH)2-D3 up-regulates both the vitamin D receptor and calbindin D-28K, whereas TPA down-regulates the expression of both proteins. These data support the involvement of PKC in the mechanism of action of 1,25-(OH)2-D3 and specifically implicate PKCβ in 1,25-(OH)2- D3-mediated nuclear events.",

author = "Maura Simboli-Campbell and Gagnon, \{Anne Marie\} and Franks, \{Douglas J.\} and Welsh, \{Jo Ellen\}",

year = "1994",

month = feb,

day = "4",

doi = "10.1016/s0021-9258(17)41856-4",

language = "English",

volume = "269",

pages = "3257--3264",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

number = "5",

}

1,25-Dihydroxyvitamin D₃ translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells. / Simboli-Campbell, Maura; Gagnon, Anne Marie; Franks, Douglas J. et al.
In: Journal of Biological Chemistry, Vol. 269, No. 5, 04.02.1994, p. 3257-3264.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - 1,25-Dihydroxyvitamin D3 translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells

AU - Simboli-Campbell, Maura

AU - Gagnon, Anne Marie

AU - Franks, Douglas J.

AU - Welsh, Jo Ellen

PY - 1994/2/4

Y1 - 1994/2/4

N2 - 1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) increases membrane- associated protein kinase C (PKC) activity and immunoreactivity in renal epithelial (Madin Darby bovine kidney, MDBK) cells (Simboli-Campbell, M., Franks, D. J., and Welsh, J. E. (1992) Cell Signalling 4, 99-109). We have now characterized the effects of 1,25-(OH)2-D3 on the subcellular localization of three individual isozymes by immunofluorescence and immunoblotting. Although the total amount of PKCα, PKCβ, and PKCζ are unaffected by 1,25-(OH)2-D3, this steroid hormone induces subcellular redistribution of both PKCα and PKCβ. Treatment with 1,25-(OH)2-D3 (100 nM, 24 h) enhances plasma membrane association of PKCα and induces translocation of PKCβ to the nuclear membrane. The effects of 1,25-(OH)2- D3 appear to be limited to the calcium-dependent PKC isozymes, since 1,25- (OH)2-D3 has no effect on the calcium independent isozyme, PKCζ. In contrast to rapid transient PKC translocation seen in response to agents which interact with membrane receptors to induce phospholipid hydrolysis, modulation of PKCα and PKCβ is observed after 24 h treatment with 1,25- (OH)2-D3. In MDBK cells, the phorbol ester 12-O-tetradecanoylphorbol-13- acetate (TPA) (100 nM, 24 h) down-regulates PKCα and, to a lesser extent, PKCζ, without altering their subcellular distribution. TPA also induces translocation of PKCβ to the nuclear membrane. MDBK cells treated with 1,25- (OH)2-D3, but not TPA, exhibit enhanced phosphorylation of endogenous nuclear proteins. In addition to the distinct effects of 1,25-(OH)2-D3 and TPA on PKC isozyme patterns, 1,25-(OH)2-D3 up-regulates both the vitamin D receptor and calbindin D-28K, whereas TPA down-regulates the expression of both proteins. These data support the involvement of PKC in the mechanism of action of 1,25-(OH)2-D3 and specifically implicate PKCβ in 1,25-(OH)2- D3-mediated nuclear events.

AB - 1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) increases membrane- associated protein kinase C (PKC) activity and immunoreactivity in renal epithelial (Madin Darby bovine kidney, MDBK) cells (Simboli-Campbell, M., Franks, D. J., and Welsh, J. E. (1992) Cell Signalling 4, 99-109). We have now characterized the effects of 1,25-(OH)2-D3 on the subcellular localization of three individual isozymes by immunofluorescence and immunoblotting. Although the total amount of PKCα, PKCβ, and PKCζ are unaffected by 1,25-(OH)2-D3, this steroid hormone induces subcellular redistribution of both PKCα and PKCβ. Treatment with 1,25-(OH)2-D3 (100 nM, 24 h) enhances plasma membrane association of PKCα and induces translocation of PKCβ to the nuclear membrane. The effects of 1,25-(OH)2- D3 appear to be limited to the calcium-dependent PKC isozymes, since 1,25- (OH)2-D3 has no effect on the calcium independent isozyme, PKCζ. In contrast to rapid transient PKC translocation seen in response to agents which interact with membrane receptors to induce phospholipid hydrolysis, modulation of PKCα and PKCβ is observed after 24 h treatment with 1,25- (OH)2-D3. In MDBK cells, the phorbol ester 12-O-tetradecanoylphorbol-13- acetate (TPA) (100 nM, 24 h) down-regulates PKCα and, to a lesser extent, PKCζ, without altering their subcellular distribution. TPA also induces translocation of PKCβ to the nuclear membrane. MDBK cells treated with 1,25- (OH)2-D3, but not TPA, exhibit enhanced phosphorylation of endogenous nuclear proteins. In addition to the distinct effects of 1,25-(OH)2-D3 and TPA on PKC isozyme patterns, 1,25-(OH)2-D3 up-regulates both the vitamin D receptor and calbindin D-28K, whereas TPA down-regulates the expression of both proteins. These data support the involvement of PKC in the mechanism of action of 1,25-(OH)2-D3 and specifically implicate PKCβ in 1,25-(OH)2- D3-mediated nuclear events.

UR - https://www.scopus.com/pages/publications/0028169695

U2 - 10.1016/s0021-9258(17)41856-4

DO - 10.1016/s0021-9258(17)41856-4

M3 - Article

C2 - 8106362

SN - 0021-9258

VL - 269

SP - 3257

EP - 3264

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 5

ER -

1,25-Dihydroxyvitamin D₃ translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells

Abstract

Access to Document

Other files and links

Fingerprint

Cite this