Previous Article | Next Article 
Molecular and Cellular Biology, January 2002, p. 69-77, Vol. 22, No. 1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.22.1.69-77.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Assembly of the RAG1/RAG2 Synaptic Complex
Cynthia L. Mundy, Nadja Patenge, Adam G. W. Matthews, and Marjorie A. Oettinger*
Department of Molecular Biology, Massachusetts General Hospital, and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02114
Received 26 June 2001/
Returned for modification 3 August 2001/
Accepted 21 September 2001
Assembly of antigen receptor genes by V(D)J recombination requires the site-specific recognition of two distinct DNA elements differing in the length of the spacer DNA that separates two conserved recognition motifs. Under appropriate conditions, V(D)J cleavage by the purified RAG1/RAG2 recombinase is similarly restricted. Double-strand breakage occurs only when these proteins are bound to a pair of complementary signals in a synaptic complex. We examine here the binding of the RAG proteins to signal sequences and find that the full complement of proteins required for synapsis of two signals and coupled cleavage can assemble on a single signal. This complex, composed of a dimer of RAG2 and at least a trimer of RAG1, remains inactive for double-strand break formation until a second complementary signal is provided. Thus, binding of the second signal activates the complex, possibly by inducing a conformational change. If synaptic complexes are formed similarly in vivo, one signal of a recombining pair may be the preferred site for RAG1/RAG2 assembly.
* Corresponding author. Mailing address: Department of Molecular Biology, Wellman 10, Massachusetts General Hospital, Boston, MA 02114. Phone: (617) 726-5967. Fax: (617) 726-5949. E-mail:
Oettinger{at}frodo.mgh.harvard.edu.
Molecular and Cellular Biology, January 2002, p. 69-77, Vol. 22, No. 1
0022-538X/01/$04.00+0 DOI: 10.1128/MCB.22.1.69-77.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
This article has been cited by other articles:
-
Shlyakhtenko, L. S., Gilmore, J., Kriatchko, A. N., Kumar, S., Swanson, P. C., Lyubchenko, Y. L.
(2009). Molecular Mechanism Underlying RAG1/RAG2 Synaptic Complex Formation. J. Biol. Chem.
284: 20956-20965
[Abstract]
[Full Text]
-
Kumar, S., Swanson, P. C.
(2009). Full-length RAG1 promotes contact with coding and intersignal sequences in RAG protein complexes bound to recombination signals paired in cis. Nucleic Acids Res
37: 2211-2226
[Abstract]
[Full Text]
-
Curry, J. D., Schlissel, M. S.
(2008). RAG2's non-core domain contributes to the ordered regulation of V(D)J recombination. Nucleic Acids Res
36: 5750-5762
[Abstract]
[Full Text]
-
Nishihara, T., Nagawa, F., Imai, T., Sakano, H.
(2008). RAG-Heptamer Interaction in the Synaptic Complex Is a Crucial Biochemical Checkpoint for the 12/23 Recombination Rule. J. Biol. Chem.
283: 4877-4885
[Abstract]
[Full Text]
-
Ranganath, S., Carpenter, A. C., Gleason, M., Shaw, A. C., Bassing, C. H., Alt, F. W.
(2008). Productive Coupling of Accessible V{beta}14 Segments and DJ{beta} Complexes Determines the Frequency of V{beta}14 Rearrangement. J. Immunol.
180: 2339-2346
[Abstract]
[Full Text]
-
Tang, M., Cecconi, C., Bustamante, C., Rio, D. C.
(2007). Analysis of P Element Transposase Protein-DNA Interactions during the Early Stages of Transposition. J. Biol. Chem.
282: 29002-29012
[Abstract]
[Full Text]
-
Ciubotaru, M., Kriatchko, A. N., Swanson, P. C., Bright, F. V., Schatz, D. G.
(2007). Fluorescence Resonance Energy Transfer Analysis of Recombination Signal Sequence Configuration in the RAG1/2 Synaptic Complex. Mol. Cell. Biol.
27: 4745-4758
[Abstract]
[Full Text]
-
Wu, C., Ranganath, S., Gleason, M., Woodman, B. B., Borjeson, T. M., Alt, F. W., Bassing, C. H.
(2007). Restriction of endogenous T cell antigen receptor beta rearrangements to Vbeta14 through selective recombination signal sequence modifications. Proc. Natl. Acad. Sci. USA
104: 4002-4007
[Abstract]
[Full Text]
-
Grundy, G. J., Hesse, J. E., Gellert, M.
(2007). Requirements for DNA hairpin formation by RAG1/2. Proc. Natl. Acad. Sci. USA
104: 3078-3083
[Abstract]
[Full Text]
-
Larijani, M., Zaheen, A., Frieder, D., Wang, Y., Wu, G. E., Edelmann, W., Martin, A.
(2005). Lack of MSH2 involvement differentiates V(D)J recombination from other non-homologous end joining events. Nucleic Acids Res
33: 6733-6742
[Abstract]
[Full Text]
-
Dai, Y., Wong, B., Yen, Y.-M., Oettinger, M. A., Kwon, J., Johnson, R. C.
(2005). Determinants of HMGB Proteins Required To Promote RAG1/2-Recombination Signal Sequence Complex Assembly and Catalysis during V(D)J Recombination. Mol. Cell. Biol.
25: 4413-4425
[Abstract]
[Full Text]
-
Auge-Gouillou, C., Brillet, B., Hamelin, M.-H., Bigot, Y.
(2005). Assembly of the mariner Mos1 Synaptic Complex. Mol. Cell. Biol.
25: 2861-2870
[Abstract]
[Full Text]
-
Ciubotaru, M., Schatz, D. G.
(2004). Synapsis of Recombination Signal Sequences Located in cis and DNA Underwinding in V(D)J Recombination. Mol. Cell. Biol.
24: 8727-8744
[Abstract]
[Full Text]
-
Nagawa, F., Hirose, S., Nishizumi, H., Nishihara, T., Sakano, H.
(2004). Joining Mutants of RAG1 and RAG2 that Demonstrate Impaired Interactions with the Coding-end DNA. J. Biol. Chem.
279: 38360-38368
[Abstract]
[Full Text]
-
Talukder, S. R., Dudley, D. D., Alt, F. W., Takahama, Y., Akamatsu, Y.
(2004). Increased frequency of aberrant V(D)J recombination products in core RAG-expressing mice. Nucleic Acids Res
32: 4539-4549
[Abstract]
[Full Text]
-
De, P., Peak, M. M., Rodgers, K. K.
(2004). DNA Cleavage Activity of the V(D)J Recombination Protein RAG1 Is Autoregulated. Mol. Cell. Biol.
24: 6850-6860
[Abstract]
[Full Text]
-
Ason, B., Reznikoff, W. S.
(2004). A high-throughput assay for Tn5 Tnp-induced DNA cleavage. Nucleic Acids Res
32: e83-e83
[Abstract]
[Full Text]
-
Perkins, E. J., Kee, B. L., Ramsden, D. A.
(2004). Histone 3 lysine 4 methylation during the pre-B to immature B-cell transition. Nucleic Acids Res
32: 1942-1947
[Abstract]
[Full Text]
-
Ko, J. E., Kim, C. W., Kim, D. R.
(2004). Amino Acid Residues in RAG1 Responsible for the Interaction with RAG2 during the V(D)J Recombination Process. J. Biol. Chem.
279: 7715-7720
[Abstract]
[Full Text]
-
Hughes, M. M., Tillman, R. E., Wehrly, T. D., White, J. M., Sleckman, B. P.
(2003). The B12/23 Restriction Is Critically Dependent on Recombination Signal Nonamer and Spacer Sequences. J. Immunol.
171: 6604-6610
[Abstract]
[Full Text]
-
Montalbano, A., Ogwaro, K. M., Tang, A., Matthews, A. G. W., Larijani, M., Oettinger, M. A., Feeney, A. J.
(2003). V(D)J Recombination Frequencies Can Be Profoundly Affected by Changes in the Spacer Sequence. J. Immunol.
171: 5296-5304
[Abstract]
[Full Text]
-
Tsai, C.-L., Chatterji, M., Schatz, D. G.
(2003). DNA mismatches and GC-rich motifs target transposition by the RAG1/RAG2 transposase. Nucleic Acids Res
31: 6180-6190
[Abstract]
[Full Text]
-
Olaru, A., Patterson, D. N., Villey, I., Livak, F.
(2003). DNA-Rag Protein Interactions in the Control of Selective D Gene Utilization in the TCR{beta} Locus. J. Immunol.
171: 3605-3611
[Abstract]
[Full Text]
-
Mathieu, N., Spicuglia, S., Gorbatch, S., Cabaud, O., Fernex, C., Verthuy, C., Hempel, W. M., Hueber, A.-O., Ferrier, P.
(2003). Assessing the Role of the T Cell Receptor beta Gene Enhancer in Regulating Coding Joint Formation during V(D)J Recombination. J. Biol. Chem.
278: 18101-18109
[Abstract]
[Full Text]
-
Godderz, L. J., Rahman, N. S., Risinger, G. M., Arbuckle, J. L., Rodgers, K. K.
(2003). Self-association and conformational properties of RAG1: implications for formation of the V(D)J recombinase. Nucleic Acids Res
31: 2014-2023
[Abstract]
[Full Text]
-
Yurchenko, V., Xue, Z., Sadofsky, M.
(2003). The RAG1 N-terminal domain is an E3 ubiquitin ligase. Genes Dev.
17: 581-585
[Abstract]
[Full Text]
-
Ciubotaru, M., Ptaszek, L. M., Baker, G. A., Baker, S. N., Bright, F. V., Schatz, D. G.
(2003). RAG1-DNA Binding in V(D)J Recombination. SPECIFICITY AND DNA-INDUCED CONFORMATIONAL CHANGES REVEALED BY FLUORESCENCE AND CD SPECTROSCOPY. J. Biol. Chem.
278: 5584-5596
[Abstract]
[Full Text]
-
Akamatsu, Y., Monroe, R., Dudley, D. D., Elkin, S. K., Gartner, F., Talukder, S. R., Takahama, Y., Alt, F. W., Bassing, C. H., Oettinger, M. A.
(2003). Deletion of the RAG2 C terminus leads to impaired lymphoid development in mice. Proc. Natl. Acad. Sci. USA
100: 1209-1214
[Abstract]
[Full Text]
-
Tillman, R. E., Wooley, A. L., Khor, B., Wehrly, T. D., Little, C. A., Sleckman, B. P.
(2003). Cutting Edge: Targeting of V{beta} to D{beta} Rearrangement by RSSs Can Be Mediated by the V(D)J Recombinase in the Absence of Additional Lymphoid-Specific Factors. J. Immunol.
170: 5-9
[Abstract]
[Full Text]
-
Swanson, P. C.
(2002). A RAG-1/RAG-2 Tetramer Supports 12/23-Regulated Synapsis, Cleavage, and Transposition of V(D)J Recombination Signals. Mol. Cell. Biol.
22: 7790-7801
[Abstract]
[Full Text]
-
Nagawa, F., Kodama, M., Nishihara, T., Ishiguro, K.-i., Sakano, H.
(2002). Footprint Analysis of Recombination Signal Sequences in the 12/23 Synaptic Complex of V(D)J Recombination. Mol. Cell. Biol.
22: 7217-7225
[Abstract]
[Full Text]
-
Tsai, C.-L., Drejer, A. H., Schatz, D. G.
(2002). Evidence of a critical architectural function for the RAG proteins in end processing, protection, and joining in V(D)J recombination. Genes Dev.
16: 1934-1949
[Abstract]
[Full Text]