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Related Experiment Videos

A specific DIF binding protein in Dictyostelium.

R Insall1, R R Kay

  • 1MRC Laboratory of Molecular Biology, Cambridge, UK.

The EMBO Journal
|October 1, 1990
PubMed
Summary

Researchers identified a DIF-1 receptor in Dictyostelium, crucial for stalk cell differentiation. This receptor

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Area of Science:

  • Cellular and Molecular Biology
  • Developmental Biology
  • Biochemistry

Background:

  • Differentiation Inducing Factor (DIF-1) regulates stalk cell differentiation in Dictyostelium.
  • The precise mechanism and receptor for DIF-1 have remained elusive.

Purpose of the Study:

  • To identify and characterize the putative DIF-1 receptor.
  • To understand the role of DIF-1 binding in Dictyostelium development.

Main Methods:

  • Cell lysate preparation and protease sensitivity assay.
  • Tritiated DIF-1 binding assays using Scatchard and linear subtraction plots.
  • Heparin-agarose chromatography.
  • Competition assays with DIF-1 analogues.
  • Developmental expression analysis and cyclic AMP induction studies.

Main Results:

  • A protease-sensitive activity binding DIF-1 was identified.
  • High-affinity (Kd = 1.8 nM) and low-abundance (1100 sites/cell) binding sites were characterized.
  • The binding activity eluted as a single peak from heparin-agarose.
  • Binding activity correlated with DIF-1 analogue potency and was developmentally regulated.
  • Cyclic AMP induced the expression of the binding activity.

Conclusions:

  • A high-affinity DIF-1 receptor exists in Dictyostelium.
  • Receptor expression is temporally regulated during development, preceding DIF induction.
  • This receptor likely plays a key role in mediating DIF-1's effects on stalk cell differentiation.

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