Identification of β-actin sequences necessary for induction by phorbol esters and calcium ionophores

Academic Article

Abstract

  • Transcription of the cytoskeletal β-actin gene is rapidly induced by phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore, A23187, in cultured H4IIE hepatoma (H4) cells. PMA directly activates protein kinase C (PKC) and activation of PKC is necessary for the cellular actions of PMA, including induction of β-actin gene transcription. In the present study, we determined the DNA sequence requirements for induction of the β-actin gene by PMA and A23187. Constructs containing progressive deletions of normal and mutated human β-actin 5' sequences fused to the reporter gene, bacterial chloramphenicol acetyltransferase, were analysed in transient transfections of H4 cells. We delineated the PMA response DNA element of the human,B-actin gene to the proximal CCArGG box (-62 to -53) in the 5' flanking region. In contrast, A23187 did not induce expression of transfected gene constructs containing this CCArGG box. Additionally, we demonstrated that CCArGG boxes from two other PMA-induced genes in H4 cells, c-fos and γ-actin, could confer PMA inducibility to a heterologous promoter. This CCArGG box specifically interacts with one or more proteins present in nuclear extracts of H4 cells. These results indicate that in cultured cells, PMA-dependent induction of the β-actin gene is mediated through the proximal CCArGG box. This suggests that the CCArGG box is a target for PKC action and may be involved in the control of other PKC regulated genes.
  • Published In

  • Oncogene  Journal
  • Author List

  • Onyia JE; Halladay DL; Messina JL
  • Start Page

  • 1713
  • End Page

  • 1722
  • Volume

  • 9
  • Issue

  • 6