Related Experiment Video
Updated: Aug 9, 2026
11:40
Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
Published on: June 25, 2013
The RdeA-RegA system, a eukaryotic phospho-relay controlling cAMP breakdown
P A Thomason1, D Traynor, J B Stock
1Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, United Kingdom. thomason@mrc-lmb.cam.ac.uk
The Journal of Biological Chemistry
|September 17, 1999
Summary
Dictyostelium development relies on cAMP signaling regulated by RegA and RdeA proteins. Genetic and biochemical evidence shows RdeA transfers phosphates to RegA, activating its phosphodiesterase activity for proper development.
Area of Science:
- Cellular signaling pathways
- Molecular mechanisms of development
- Eukaryotic signal transduction
Background:
- The regA and rdeA genes in Dictyostelium encode proteins crucial for regulating cyclic AMP (cAMP) signaling.
- RegA is a response regulator with a receiver domain and a cAMP-phosphodiesterase domain.
- RdeA is hypothesized to be a phospho-transfer protein supplying phosphates to RegA.
Purpose of the Study:
- To investigate the functional relationship between RegA and RdeA in Dictyostelium.
- To determine the role of protein phosphorylation in RegA activity.
- To elucidate the mechanism of phospho-transfer between RdeA and RegA.
Main Methods:
- Genetic analysis in Dictyostelium.
- Biochemical assays for protein-protein interaction and phospho-transfer.
- Site-directed mutagenesis to identify key amino acid residues.
Main Results:
- Phosphorylated RegA (phospho-RegA) is the active form of the enzyme in vivo, with aspartate phosphorylation at position 212 being critical.
- RdeA and RegA directly interact and transfer phosphates in vitro, dependent on histidine 65 of RdeA and aspartate 212 of RegA.
- Phosphorylation by RdeA or a heterologous donor significantly enhances RegA's phosphodiesterase activity (at least 20-fold).
Conclusions:
- RegA and RdeA form a eukaryotic His-Asp phospho-relay system regulating Dictyostelium development.
- This signaling network is essential for Dictyostelium development and fruiting body maturation.
- The findings reveal a novel mechanism for signal transduction in eukaryotes.
More Related Videos
09:04Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
Published on: July 26, 2018
09:26DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
Published on: December 29, 2021
Related Concept Videos
01:23Negative Regulator Molecules
Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
02:36DNA Damage can Stall the Cell Cycle
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
02:31Homologous Recombination
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
02:37Restarting Stalled Replication Forks
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
02:36DNA Damage Can Stall the Cell Cycle
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
01:28The Cell Cycle Control System
The cell cycle regulation directs how a cell proceeds from one phase to the next and begins mitosis. The cell cycle control system includes intracellular regulatory molecules and external triggers. They provide "stop" or "advance" signals and operate at specific cell cycle stages termed checkpoints to ensure that a particular process is completed before the cell advances to the next phase.
Cyclins and cyclin-dependent kinases (Cdks) are the primary cell cycle regulators and function at the cell...
Cyclins and cyclin-dependent kinases (Cdks) are the primary cell cycle regulators and function at the cell...