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J. Biol. Chem., Vol. 269, Issue 51, 32233-32238, 12, 1994
JP Berg, AH Ree, JA Sandvik, K Tasken, BF Landmark, PA Torjesen and E Haug
1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) attenuates the stimulatory effects
of cAMP on proliferation and iodide uptake in rat thyroid FRTL- 5 cells.
This study examines the effects of 1,25-(OH)2D3 on the cAMP- dependent
protein kinase (PKA). Cytosol proteins separated by anion exchange
chromatography showed increased [3H]cAMP binding activity as well as
increased kinase activity in the fractions containing PKA type II in
1,25-(OH)2D3 (10 nM)-treated cells compared to the control cells. Western
blot analysis of 1,25-(OH)2D3-treated cells revealed a 4-fold increase in
the cytosolic amount of the PKA regulatory subunit RII beta, whereas no
changes were detected in the regulatory subunits RI alpha and RII alpha or
the catalytic (C) subunit. Northern blot analyses showed a similar increase
in RII beta mRNA in cells treated for 12 h with 1,25-(OH)2D3 (10 nM), and
RII beta mRNA increased further to 10-fold above control cell level after
96 h of incubation. Iodide uptake was synergistically stimulated with both
PKAI- and PKAII- directed pairs of cAMP analogs. The PKAI synergism was,
however, inhibited by 1,25-(OH)2D3 treatment of the cells, whereas the
PKAII synergism was unaffected. In conclusion, 1,25-(OH)2D3 attenuates both
PKAI formation and PKAI-stimulated iodide uptake in rat thyroid FRTL-5
cells by increasing the level of RII beta without altering the other PKA
subunit levels.
1,25-dihydroxyvitamin D3 alters the effect of cAMP in thyroid cells by increasing the regulatory subunit type II beta of the cAMP-dependent protein kinase
Hormone Laboratory, Aker Hospital, Oslo, Norway.
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