Cited by 47DNA Repair MechanismsT-cell and B-cell ImmunologyGenomics and Chromatin Dynamics

Definition of Minimal Domains of Interaction Within the Recombination-Activating Genes 1 and 2 Recombinase Complex

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Aidinis, Vassilis, Dias, Dora C, Gomez, Carlos A, Bhattacharyya, Debika, Spanopoulou, Eugenia, Santagata, Sandro·2000·The Journal of Immunology

Abstract

During V(D)J recombination, recognition and cleavage of the recombination signal sequences (RSSs) requires the coordinated action of the recombination-activating genes 1 and 2 (RAG1/RAG2) recombinase complex. In this report, we use deletion mapping and site-directed mutagenesis to determine the minimal domains critical for interaction between RAG1 and RAG2. We define the active core of RAG2 required for RSS cleavage as aa 1–371 and demonstrate that the C-terminal 57 aa of this core provide a dominant surface for RAG1 interaction. This region corresponds to the last of six predicted kelch repeat motifs that have been proposed by sequence analysis to fold RAG2 into a six-bladed β-propeller structure. Residue W317 within this sixth repeat is shown to be critical for mediating contact with RAG1 and concurrently for stabilizing binding and directing cleavage of the RSS. We also show that zinc finger B (aa 727–750) of RAG1 provides a dominant interaction domain for recruiting RAG2. In all, the data support a model of RAG2 as a multimodular protein that utilizes one of its six faces for establishing productive contacts with RAG1.

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  • Only two cysteines of the zinc finger B (ZFB) C2H2 motif in RAG1 (residues 695-761) involved in its binding to RAG2 are perfectly conserved in the Transib TPases (motif 7).

    “Interestingly, only two cysteines of the zinc finger B (ZFB) C2H2 motif in RAG1 (residues 695–761) involved in its binding to RAG2 [30,31] are perfectly conserved in the Transib TPases (motif 7; see Figures 3 and Figure S3).”

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Authors
Aidinis, V., Dias, D. C., Gomez, C. A., Bhattacharyya, D., Spanopoulou, E., & Santagata, S.
Affiliations
Icahn School of Medicine at Mount Sinai (US)
Venue
The Journal of Immunology
Published
2000-06-01
Volume
164
Issue
11
Pages
5826-5832
ISSN
  • 0022-1767 (ISSN-L)
  • 1550-6606
OpenAlex
W1580095468
Cited by
47
References
62
Citations by year
2025: 1 · 2024: 2 · 2023: 2 · 2020: 1 · 2016: 1 · 2015: 4 · 2014: 3 · 2013: 1
FWCI
1.81
Type
journal article
Language
en
Open access
Locations
  • The Journal of Immunology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Cancer Research Institute
  • National Institutes of Health (5r01ai040191-04)
  • National Institute of Allergy and Infectious Diseases (AI40191)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
RAG1, RAG2, V(D)J recombination, protein-protein interaction domains, kelch repeats, six-bladed β-propeller, immunoglobulin gene rearrangement
MeSH terms
Amino Acid Sequence, Cell Line, DNA Nucleotidyltransferases, DNA-Binding Proteins, Humans, Molecular Sequence Data, Nuclear Proteins, Peptide Fragments, Chemical Precipitation, Tryptophan, Mutagenesis, Site-Directed, Zinc Fingers, Genes, RAG-1, Protein Structure, Tertiary, Homeodomain Proteins, Amino Acid Substitution, VDJ Recombinases

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