Sp1 transcriptional activity is up-regulated by phosphatase 2A in dividing T lymphocytes.
We have followed Sp1 expression in primary human T lymphocytes induced, via CD2 plus CD28 costimulation, to sustained proliferation and subsequent return to quiescence. Binding of Sp1 to wheat germ agglutinin lectin was not modified following activation, indicating that the overall glycosylation of the protein was unchanged. Sp1 underwent, instead, a major dephosphorylation that correlated with cyclin A expression and, thus, with cell cycle progression. A similar change was observed in T cells that re-entered cell cycle following secondary interleukin-2 stimulation, as well as in serum-induced proliferating NIH/3T3 fibroblasts. Phosphatase 2A (PP2A) appears involved because 1) treatment of dividing cells with okadaic acid or cantharidin inhibited Sp1 dephosphorylation and 2) PP2A dephosphorylated Sp1 in vitro and strongly interacted with Sp1 in vivo. Sp1 dephosphorylation is likely to increase its transcriptional activity because PP2A overexpression potentiated Sp1 site-driven chloramphenicol acetyltransferase expression in dividing Kit225 T cells and okadaic acid reversed this effect. This increase might be mediated by a stronger affinity of dephosphorylated Sp1 for DNA, as illustrated by the reduced DNA occupancy by hyperphosphorylated Sp factors from cantharidin- or nocodazole-treated cells. Finally, Sp1 dephosphorylation appears to occur throughout cell cycle except for mitosis, a likely common feature to all cycling cells.
Lacroix
Isabelle
I
Unité de Cancérologie Expérimentale, U119 INSERM, 27 boulevard Lei Roure, 13009 Marseille, France.
Lipcey
Carol
C
Imbert
Jean
J
Kahn-Perlès
Brigitte
B
D002453:Cell Cycle; D004247:DNA / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008938:Mitosis; D010749:Phosphoprotein Phosphatases / Q000502:physiology*; D010766:Phosphorylation; D054648:Protein Phosphatase 2; D016329:Sp1 Transcription Factor / Q000096:biosynthesis*; D013601:T-Lymphocytes / Q000378:metabolism*; D015854:Up-Regulation
D007376:Interleukin-2; D016329:Sp1 Transcription Factor; D004247:DNA; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2