Expression of the contact site A glycoprotein in Dictyostelium discoideum: quantitation and developmental regulation

Abstract

We have previously reported that a monoclonal antibody directed specifically against a surface glycoprotein of Mr 80,000 (gp80) inhibits the EDTA-resistant contact sites A of Dictyostelium discoideum (Siu, C.-H., and Choi, A.H.C. (1985) J. Biol. Chem. 260, 16030-16036). In this report, we describe an assay using this monoclonal antibody to quantitate the amount of gp80 expressed at different developmental stages. Under normal conditions, gp80 is detectable after 6 h of development and it rapidly accumulates between 6 and 10 h, corresponding to the time when cells acquire their EDTA-resistant binding sites. At the peak level, there are 1.5.10(5) gp80 molecules per cell. More than 90% of the cellular gp80 is located on the cell surface. When cells are given exogenous pulses of cAMP, a precocious and enhanced expression of gp80 is induced. At the peak level, the cAMP-pulsed cells accumulate five times more gp80 than the non-pulsed cells. This is preceeded by an equally rapid accumulation of gp80 transcripts, suggesting that cAMP regulates gp80 synthesis at the transcriptional level.