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Transmembrane Domain Oligomerization Propensity determined by ToxR Assay
Published on: May 26, 2011
Thermal dependency of RAG1 self-association properties
Pallabi De1, Shuying Zhao, Lori M Gwyn
1Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190, USA. pallabi.de@umassmed.edu
BMC Biochemistry
|February 1, 2008
Summary
The RAG1 protein functions as a dimer in V(D)J recombination. Its stability and interaction with RAG2 and the recombination signal sequence (RSS) are crucial for lymphocyte development.
Area of Science:
- Molecular biology
- Immunology
- Biochemistry
Background:
- V(D)J recombination generates functional immunoglobulin and T cell receptor genes in developing lymphocytes.
- The V(D)J recombinase, comprising RAG1 and RAG2, catalyzes DNA cleavage at specific sites marked by recombination signal sequences (RSS).
- RAG1 possesses the active site and RSS binding domains, while RAG2 is essential for DNA cleavage, yet their physicochemical properties and interactions are not fully understood.
Purpose of the Study:
- To investigate the self-association properties of the RAG1 protein.
- To determine the oligomeric state of RAG1 required for interaction with RAG2 and the RSS.
- To elucidate the role of RAG2 in stabilizing the RAG1-RSS complex under physiological conditions.
Main Methods:
- Analysis of RAG1 self-association properties.
- Assessment of RAG1 oligomer interactions with RAG2 and RSS.
- Investigation of complex formation at physiological temperature (37°C).
Main Results:
- RAG1 exists in multiple oligomeric forms, but only the dimeric form interacts with RAG2 and the RSS.
- Dimeric RAG1 can aggregate and become inactive at physiological temperatures without RAG2.
- The addition of RAG2 stabilizes the V(D)J recombinase:RSS complex, which contains a single RAG1 dimer at 37°C.
Conclusions:
- The functional form of RAG1 in V(D)J recombination is dimeric.
- Complex formation with RAG2 is likely necessary for RAG1 stability under physiological conditions.
- Temperature-dependent self-association of RAG1 must be considered in future structural and functional studies.
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