Differential involvement of the actin cytoskeleton in differentiation and mitogenesis of thyroid cells: inactivation of Rho proteins contributes to cyclic adenosine monophosphate-dependent gene expression but prevents mitogenesis.
In thyroid epithelial cells, TSH via cAMP induces a rounding up of the cells associated with actin stress fiber disruption, expression of differentiation genes and cell cycle progression. Here we have evaluated the role of small G proteins of the Rho family and their impact on the actin cytoskeleton in these different processes in primary cultures of canine thyrocytes. TSH and forskolin, but not growth factors, rapidly inactivated RhoA, Rac1, and Cdc42, as assayed by detection of GTP-bound forms. Using toxins that inactivate Rho proteins (toxin B, C3 exoenzyme) or activate them [cytotoxic necrotizing factor 1 (CNF1)], in comparison with disruption of the actin cytoskeleton by dihydrocytochalasin B (DCB) or latrunculin, two unexpected conclusions were reached: 1) inactivation of Rho proteins by cAMP, by disorganizing actin microfilaments and inducing cell retraction, could be necessary and sufficient to mediate at least part of the cAMP-dependent induction of thyroglobulin and thyroid oxidases, but only partly necessary for the induction of Na(+)/I(-) symporter and thyroperoxidase; 2) as indicated by the effect of their inhibition by toxin B and C3, some residual activity of Rho proteins could be required for the induction by cAMP-dependent or -independent mitogenic cascades of DNA synthesis and retinoblastoma protein (pRb) phosphorylation, through mechanisms targeting the activity, but not the stimulated assembly, of cyclin D3-cyclin-dependent kinase 4 complexes. However, at variance with current concepts mostly derived from fibroblast models, DNA synthesis induction and cyclin D3-cyclin-dependent kinase 4 activation were resistant to actin depolymerization by dihydrocytochalasin B in canine thyrocytes, which provides a first such example in a normal adherent cell.
Fortemaison
Nathalie
N
Institute of Interdisciplinary Research, Université Libre de Bruxelles, Belgium.
Blancquaert
Sara
S
Dumont
Jacques E
JE
Maenhaut
Carine
C
Aktories
Klaus
K
Roger
Pierre P
PP
Dremier
Sarah
S
D000199:Actins / Q000502:physiology*; D000209:Acute-Phase Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology*; D004285:Dogs; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology*; D006133:Growth Substances / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects / Q000502:physiology; D013961:Thyroid Gland / Q000166:cytology* / Q000187:drug effects; D013972:Thyrotropin / Q000494:pharmacology; D014118:Toxins, Biological / Q000494:pharmacology; D020764:cdc42 GTP-Binding Protein / Q000037:antagonists & inhibitors; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors
D000199:Actins; D000209:Acute-Phase Proteins; D006133:Growth Substances; D014118:Toxins, Biological; C010781:acute-phase protein rho; D013972:Thyrotropin; D000242:Cyclic AMP; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein