Synthesis of the Ca(2+)-dependent cell adhesion molecule DdCAD-1 is regulated by multiple factors during Dictyostelium development.
In Dictyostelium discoideum, the cadA gene encodes the cell adhesion molecule DdCAD-1, a protein of M(r) 24,000, which mediates Ca(2+)-dependent cell-cell adhesion during development. We have examined the effects of cAMP, cell-cell contact, and growth conditions on cadA expression. cadA has a unique pattern of expression, which appears to be a combination of the expression patterns of early genes and aggregation-stage genes. Expression of the cadA gene in bacterially grown cells is activated at the beginning of the developmental cycle, followed by a period of rapid DdCAD-1 accumulation. The mRNA level reaches its maximum at 9 h of development and then declines to the basal level at approximately 18 h, while the protein level remains constant after reaching its maximum at 12 h. Pulse-chase experiments have demonstrated that DdCAD-1 has a significantly longer half-life than the average cellular protein. Transcription of the cadA gene is stimulated by exogenous cAMP pulses, leading to a 3- to 5-fold increase in the transcription rate. In the fgdA mutant, which lacks a functional G alpha 2, cAMP fails to enhance cadA expression, suggesting that cAMP stimulates cadA transcription via a G protein-dependent pathway. However, inhibition of cell-cell contact has no effect on the synthesis of DdCAD-1. Growth conditions also have a major influence on cadA expression. Axenically grown cells produce a high level of cadA transcripts during vegetative growth. The mRNA level shows a steady decrease during development and is reduced to the basal level by 12 h. In contrast, the level of DdCAD-1 remains relatively high throughout development, suggesting that axenic growth affects the accumulation of cadA mRNA but not the stability of the protein. These results indicate that multiple mechanisms are involved to maintain a high level of DdCAD-1 during development.
Yang
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C
Banting and Best Department of Medical Research, University of Toronto, Ontario, Canada.
Brar
S K
SK
Desbarats
L
L
Siu
C H
CH
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000235:genetics; D002450:Cell Communication / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000254:growth & development / Q000378:metabolism*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D009154:Mutation
D002135:Calcium-Binding Proteins; D015815:Cell Adhesion Molecules; D003470:Culture Media; C100604:DdCAD-1 protein, Dictyostelium discoideum; D005656:Fungal Proteins; D000242:Cyclic AMP; D002118:Calcium