A temperature-sensitive adenylyl cyclase mutant of Dictyostelium

Hitesh Patel, Kunde Guo, Carole Parent, Julian Gross, Peter N. Devreotes, Cornells J. Weijer

Research output: Contribution to journalArticlepeer-review

Abstract

Dictyostelium development starts with the chemotactic aggregation of up to 106 amoebae in response to propagating cAMP waves. cAMP is produced by the aggregation stage adenylyl cyclase (ACA) and cells lacking ACA (aca null) cannot aggregate. Temperature-sensitive mutants of ACA were selected from a population of aca null cells transformed with a library of ACA genes, a major segment of which had been amplified by error-prone PCR. One mutant (tsaca2) that can complement the aggregation null phenotype of aca null cells at 22°C but not at 28°C was characterized in detail. The basal catalytic activity of the enzyme in this mutant was rapidly and reversibly inactivated at 28°C. Using this mutant strain we show that cell movement in aggregates and mounds is organized by propagating waves of cAMP. Synergy experiments between wild-type and tsaca2 cells, shifted to the restrictive temperature at various stages of development, showed that ACA plays an important role in the control of cell sorting and tip formation.

Original languageEnglish (US)
Pages (from-to)2247-2256
Number of pages10
JournalEMBO Journal
Volume19
Issue number10
DOIs
StatePublished - May 15 2000

Keywords

  • Adenylyl cyclase
  • Cell movement
  • Cell sorting
  • Differentiation
  • cAMP relay chemotaxis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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