Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 15 March 1996:
Vol. 271. no. 5255, pp. 1592 - 1594
DOI: 10.1126/science.271.5255.1592
Prev | Table of Contents | Next

Reports

Similarities Between Initiation of V(D)J Recombination and Retroviral Integration

Dik C. van Gent, Kiyoshi Mizuuchi, Martin Gellert *

In the first step of V(D)J recombination, the RAG1 and RAG2 proteins cleave DNA between a signal sequence and the adjacent coding sequence, generating a blunt signal end and a coding end with a closed hairpin structure. These hairpins are intermediates leading to the formation of assembled antigen receptor genes. It is shown here that the hairpins are formed by a chemical mechanism of direct trans-esterification, very similar to the early steps of transpositional recombination and retroviral integration. A minor variation in the reaction is sufficient to divert the process from transposition to hairpin formation.

Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0540, USA.
* To whom correspondence should be addressed.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Resistance Mutations in Human Immunodeficiency Virus Type 1 Integrase Selected with Elvitegravir Confer Reduced Susceptibility to a Wide Range of Integrase Inhibitors.
O. Goethals, R. Clayton, M. Van Ginderen, I. Vereycken, E. Wagemans, P. Geluykens, K. Dockx, R. Strijbos, V. Smits, A. Vos, et al. (2008)
J. Virol. 82, 10366-10374
   Abstract »    Full Text »    PDF »
RAG-Heptamer Interaction in the Synaptic Complex Is a Crucial Biochemical Checkpoint for the 12/23 Recombination Rule.
T. Nishihara, F. Nagawa, T. Imai, and H. Sakano (2008)
J. Biol. Chem. 283, 4877-4885
   Abstract »    Full Text »    PDF »
G.O.D.'s Holy Grail: Discovery of the RAG Proteins.
V. L. Brandt and D. B. Roth (2008)
J. Immunol. 180, 3-4
   Full Text »    PDF »
The Beyond 12/23 Restriction Is Imposed at the Nicking and Pairing Steps of DNA Cleavage during V(D)J Recombination.
A. H. Drejer-Teel, S. D. Fugmann, and D. G. Schatz (2007)
Mol. Cell. Biol. 27, 6288-6299
   Abstract »    Full Text »    PDF »
Control of transposase activity within a transpososome by the configuration of the flanking DNA segment of the transposon.
M. Mizuuchi, P. A. Rice, S. J. Wardle, D. B. Haniford, and K. Mizuuchi (2007)
PNAS 104, 14622-14627
   Abstract »    Full Text »    PDF »
DNA transposition target immunity and the determinants of the MuB distribution patterns on DNA.
X. Tan, M. Mizuuchi, and K. Mizuuchi (2007)
PNAS 104, 13925-13929
   Abstract »    Full Text »    PDF »
Site-specific DNA transesterification catalyzed by a restriction enzyme.
G. Sasnauskas, B. A. Connolly, S. E. Halford, and V. Siksnys (2007)
PNAS 104, 2115-2120
   Abstract »    Full Text »    PDF »
Identification and Characterization of a Gain-of-Function RAG-1 Mutant.
A. N. Kriatchko, D. K. Anderson, and P. C. Swanson (2006)
Mol. Cell. Biol. 26, 4712-4728
   Abstract »    Full Text »    PDF »
Genomic instability due to V(D)J recombination-associated transposition..
Y. V.R. Reddy, E. J. Perkins, and D. A. Ramsden (2006)
Genes & Dev. 20, 1575-1582
   Abstract »    Full Text »    PDF »
Both High Mobility Group (HMG)-boxes and the Acidic Tail of HMGB1 Regulate Recombination-activating Gene (RAG)-mediated Recombination Signal Synapsis and Cleavage in Vitro.
S. Bergeron, T. Madathiparambil, and P. C. Swanson (2005)
J. Biol. Chem. 280, 31314-31324
   Abstract »    Full Text »    PDF »
The MRE11-RAD50-XRS2 Complex, in Addition to Other Non-homologous End-joining Factors, Is Required for V(D)J Joining in Yeast.
A. E. Clatworthy, M. A. Valencia-Burton, J. E. Haber, and M. A. Oettinger (2005)
J. Biol. Chem. 280, 20247-20252
   Abstract »    Full Text »    PDF »
Targeting Tn5 Transposase Identifies Human Immunodeficiency Virus Type 1 Inhibitors.
B. Ason, D. J. Knauss, A. M. Balke, G. Merkel, A. M. Skalka, and W. S. Reznikoff (2005)
Antimicrob. Agents Chemother. 49, 2035-2043
   Abstract »    Full Text »    PDF »
Cyclic changes in the affinity of protein-DNA interactions drive the progression and regulate the outcome of the Tn10 transposition reaction.
D. Liu, P. Crellin, and R. Chalmers (2005)
Nucleic Acids Res. 33, 1982-1992
   Abstract »    Full Text »    PDF »
Histone H2AX Is Phosphorylated at Sites of Retroviral DNA Integration but Is Dispensable for Postintegration Repair.
R. Daniel, J. Ramcharan, E. Rogakou, K. D. Taganov, J. G. Greger, W. Bonner, A. Nussenzweig, R. A. Katz, and A. M. Skalka (2004)
J. Biol. Chem. 279, 45810-45814
   Abstract »    Full Text »    PDF »
In Vitro Processing of the 3'-Overhanging DNA in the Postcleavage Complex Involved in V(D)J Joining.
T. Nishihara, F. Nagawa, H. Nishizumi, M. Kodama, S. Hirose, R. Hayashi, and H. Sakano (2004)
Mol. Cell. Biol. 24, 3692-3702
   Abstract »    Full Text »    PDF »
Full-length RAG-2, and Not Full-length RAG-1, Specifically Suppresses RAG-mediated Transposition but Not Hybrid Joint Formation or Disintegration.
P. C. Swanson, D. Volkmer, and L. Wang (2004)
J. Biol. Chem. 279, 4034-4044
   Abstract »    Full Text »    PDF »
Site-specific recombination by the DDE family member mobile element IS30 transposase.
J. Kiss, M. Szabo, and F. Olasz (2003)
PNAS 100, 15000-15005
   Abstract »    Full Text »    PDF »
A RAG-1/RAG-2 Tetramer Supports 12/23-Regulated Synapsis, Cleavage, and Transposition of V(D)J Recombination Signals.
P. C. Swanson (2002)
Mol. Cell. Biol. 22, 7790-7801
   Abstract »    Full Text »    PDF »
Fine Structure and Activity of Discrete RAG-HMG Complexes on V(D)J Recombination Signals.
P. C. Swanson (2002)
Mol. Cell. Biol. 22, 1340-1351
   Abstract »    Full Text »    PDF »
Assembly of the RAG1/RAG2 Synaptic Complex.
C. L. Mundy, N. Patenge, A. G. W. Matthews, and M. A. Oettinger (2002)
Mol. Cell. Biol. 22, 69-77
   Abstract »    Full Text »    PDF »
Effect of HIV integrase inhibitors on the RAG1/2 recombinase.
M. Melek, J. M. Jones, M. H. O'Dea, G. Pais, T. R. Burke Jr., Y. Pommier, N. Neamati, and M. Gellert (2001)
PNAS
   Abstract »    Full Text »    PDF »
The Role of Recombination Activating Gene (RAG) Reinduction in Thymocyte Development in Vivo.
N. Yannoutsos, P. Wilson, W. Yu, H. T. Chen, A. Nussenzweig, H. Petrie, and M. C. Nussenzweig (2001)
J. Exp. Med. 194, 471-480
   Abstract »    Full Text »    PDF »
RAG-1 Mutations Associated with B-Cell-Negative SCID Dissociate the Nicking and Transesterification Steps of V(D)J Recombination.
W. Li, F.-C. Chang, and S. Desiderio (2001)
Mol. Cell. Biol. 21, 3935-3946
   Abstract »    Full Text »    PDF »
Regulation of Activator/Dissociation Transposition by Replication and DNA Methylation.
F. Ros and R. Kunze (2001)
Genetics 157, 1723-1733
   Abstract »    Full Text »
The DDE Motif in RAG-1 Is Contributed in trans to a Single Active Site That Catalyzes the Nicking and Transesterification Steps of V(D)J Recombination.
P. C. Swanson (2001)
Mol. Cell. Biol. 21, 449-458
   Abstract »    Full Text »    PDF »
Conditional RAG-1 Mutants Block the Hairpin Formation Step of V(D)J Recombination.
S. B. Kale, M. A. Landree, and D. B. Roth (2001)
Mol. Cell. Biol. 21, 459-466
   Abstract »    Full Text »    PDF »
Mutations in Conserved Regions of the Predicted RAG2 Kelch Repeats Block Initiation of V(D)J Recombination and Result in Primary Immunodeficiencies.
C. A. Gomez, L. M. Ptaszek, A. Villa, F. Bozzi, C. Sobacchi, E. G. Brooks, L. D. Notarangelo, E. Spanopoulou, Z. Q. Pan, P. Vezzoni, et al. (2000)
Mol. Cell. Biol. 20, 5653-5664
   Abstract »    Full Text »    PDF »
Three-Dimensional Structure of the Tn5 Synaptic Complex Transposition Intermediate.
D. R. Davies, I. Y. Goryshin, W. S. Reznikoff, and I. Rayment (2000)
Science 289, 77-85
   Abstract »    Full Text »
The Old and the Restless.
S. M. Lewis and G. E. Wu (2000)
J. Exp. Med. 191, 1631-1636
   Full Text »    PDF »
Mechanistic Basis for Coding End Sequence Effects in the Initiation of V(D)J Recombination.
K. Yu and M. R. Lieber (1999)
Mol. Cell. Biol. 19, 8094-8102
   Abstract »    Full Text »    PDF »
Mutational analysis of RAG1 and RAG2 identifies three catalytic amino acids in RAG1 critical for both cleavage steps of V(D)J recombination.
M. A. Landree, J. A. Wibbenmeyer, and D. B. Roth (1999)
Genes & Dev. 13, 3059-3069
   Abstract »    Full Text »
Mutations of acidic residues in RAG1 define the active site of the V(D)J recombinase.
D. R. Kim, Y. Dai, C. L. Mundy, W. Yang, and M. A. Oettinger (1999)
Genes & Dev. 13, 3070-3080
   Abstract »    Full Text »
Stereospecificity of Reactions Catalyzed by HIV-1 Integrase.
J. L. Gerton, D. Herschlag, and P. O. Brown (1999)
J. Biol. Chem. 274, 33480-33487
   Abstract »    Full Text »    PDF »
Involvement of DNA End-Binding Protein Ku in Ty Element Retrotransposition.
J. A. Downs and S. P. Jackson (1999)
Mol. Cell. Biol. 19, 6260-6268
   Abstract »    Full Text »    PDF »
Coordinate Regulation of RAG1 and RAG2 by Cell Type-Specific DNA Elements 5' of RAG2.
W. Yu, Z. Misulovin, H. Suh, R. R. Hardy, M. Jankovic, N. Yannoutsos, and M. C. Nussenzweig (1999)
Science 285, 1080-1084
   Abstract »    Full Text »
A RAG1 and RAG2 Tetramer Complex Is Active in Cleavage in V(D)J Recombination.
T. Bailin, X. Mo, and M. J. Sadofsky (1999)
Mol. Cell. Biol. 19, 4664-4671
   Abstract »    Full Text »    PDF »
Epigenetic Interactions among Three dTph1 Transposons in Two Homologous Chromosomes Activate a New Excision–Repair Mechanism in Petunia.
A. van Houwelingen, E. Souer, J. Mol, and R. Koes (1999)
PLANT CELL 11, 1319-1336
   Abstract »    Full Text »
DNA Hairpin Opening Mediated by the RAG1 and RAG2 Proteins.
P. E. Shockett and D. G. Schatz (1999)
Mol. Cell. Biol. 19, 4159-4166
   Abstract »    Full Text »    PDF »
RAG-2 Promotes Heptamer Occupancy by RAG-1 in the Assembly of a V(D)J Initiation Complex.
P. C. Swanson and S. Desiderio (1999)
Mol. Cell. Biol. 19, 3674-3683
   Abstract »    Full Text »    PDF »
Detection of RAG Protein-V(D)J Recombination Signal Interactions Near the Site of DNA Cleavage by UV Cross-Linking.
Q. M. Eastman, I. J. Villey, and D. G. Schatz (1999)
Mol. Cell. Biol. 19, 3788-3797
   Abstract »    Full Text »    PDF »
The Three-dimensional Structure of a Tn5 Transposase-related Protein Determined to 2.9-A Resolution.
D. R. Davies, L. M. Braam, W. S. Reznikoff, and I. Rayment (1999)
J. Biol. Chem. 274, 11904-11913
   Abstract »    Full Text »    PDF »
A Role for DNA-PK in Retroviral DNA Integration.
R. Daniel, R. A. Katz, and A. M. Skalka (1999)
Science 284, 644-647
   Abstract »    Full Text »
HMG Protein Family Members Stimulate Human Immunodeficiency Virus Type 1 and Avian Sarcoma Virus Concerted DNA Integration In Vitro.
P. Hindmarsh, T. Ridky, R. Reeves, M. Andrake, A. M. Skalka, and J. Leis (1999)
J. Virol. 73, 2994-3003
   Abstract »    Full Text »    PDF »
The Mechanism of Chromosome 7 Inversion in Human Lymphocytes Expressing Chimeric {gamma}{beta} TCR.
C. Retiere, F. Halary, M.-A. Peyrat, F. Le Deist, M. Bonneville, and M.-M. Hallet (1999)
J. Immunol. 162, 903-910
   Abstract »    Full Text »    PDF »
V(D)J Recombination: Links to Transposition and Double-strand Break Repair.
M. GELLERT, J.E. HESSE, K. HIOM, M. MELEK, M. MODESTI, T.T. PAULL, D.A. RAMSDEN, and D.C. VAN GENT (1999)
Cold Spring Harb Symp Quant Biol 64, 161-168
   Abstract »    PDF »
The RAG-HMG1 Complex Enforces the 12/23 Rule of V(D)J Recombination Specifically at the Double-Hairpin Formation Step.
R. B. West and M. R. Lieber (1998)
Mol. Cell. Biol. 18, 6408-6415
   Abstract »    Full Text »    PDF »
Pathological and Physiological Double-Strand Breaks : Roles in Cancer, Aging, and the Immune System.
M. R. Lieber (1998)
Am. J. Pathol. 153, 1323-1332
   Abstract »    Full Text »    PDF »
A Preferred Target DNA Structure for Retroviral Integrase in Vitro.
R. A. Katz, K. Gravuer, and A. M. Skalka (1998)
J. Biol. Chem. 273, 24190-24195
   Abstract »    Full Text »    PDF »
Distinct Roles of RAG1 and RAG2 in Binding the V(D)J Recombination Signal Sequences.
Y. Akamatsu and M. A. Oettinger (1998)
Mol. Cell. Biol. 18, 4670-4678
   Abstract »    Full Text »    PDF »
Functional Analysis of Coordinated Cleavage in V(D)J Recombination.
D. R. Kim and M. A. Oettinger (1998)
Mol. Cell. Biol. 18, 4679-4688
   Abstract »    Full Text »    PDF »
Extensive, Nonrandom Diversity of Excision Footprints Generated by Ds-Like Transposon Ascot-1 Suggests New Parallels with V(D)J Recombination.
V. Colot, V. Haedens, and J.-L. Rossignol (1998)
Mol. Cell. Biol. 18, 4337-4346
   Abstract »    Full Text »    PDF »
The Effect of Me2+ Cofactors at the Initial Stages of V(D)J Recombination.
S. Santagata, V. Aidinis, and E. Spanopoulou (1998)
J. Biol. Chem. 273, 16325-16331
   Abstract »    Full Text »    PDF »
Functional Characterization of the Tn5 Transposase by Limited Proteolysis.
L. A. M. Braam and W. S. Reznikoff (1998)
J. Biol. Chem. 273, 10908-10913
   Abstract »    Full Text »    PDF »
Rejoining of DNA by the RAG1 and RAG2 Proteins.
M. Melek, M. Gellert, and D. C. van Gent (1998)
Science 280, 301-303
   Abstract »    Full Text »
Structure of Nonhairpin Coding-End DNA Breaks in Cells Undergoing V(D)J Recombination.
M. S. Schlissel (1998)
Mol. Cell. Biol. 18, 2029-2037
   Abstract »    Full Text »    PDF »
Drosophila P-element transposase is a novel site-specific endonuclease.
E. L. Beall and D. C. Rio (1997)
Genes & Dev. 11, 2137-2151
   Abstract »    Full Text »    PDF »
V(D)J Recombination: Modulation of RAG1 and RAG2 Cleavage Activity on 12/23 Substrates by Whole Cell Extract and DNA-bending Proteins.
D. J. Sawchuk, F. Weis-Garcia, S. Malik, E. Besmer, M. Bustin, M. C. Nussenzweig, and P. Cortes (1997)
J. Exp. Med. 185, 2025-2032
   Abstract »    Full Text »    PDF »
Disruption of the terminal base pairs of retroviral DNA during integration..
B P Scottoline, S Chow, V Ellison, and P O Brown (1997)
Genes & Dev. 11, 371-382
   Abstract »    PDF »
Retroviral Integrase, Putting the Pieces Together.
M. D. Andrake and A. M. Skalka (1996)
J. Biol. Chem. 271, 19633-19636
   Full Text »    PDF »
Degradation of HIV-1 Integrase by the N-end Rule Pathway.
L. C. F. Mulder and M. A. Muesing (2000)
J. Biol. Chem. 275, 29749-29753
   Abstract »    Full Text »    PDF »
Identification of Two Topologically Independent Domains in RAG1 and Their Role in Macromolecular Interactions Relevant to V(D)J Recombination.
J. L. Arbuckle, L. J. Fauss, R. Simpson, L. M. Ptaszek, and K. K. Rodgers (2001)
J. Biol. Chem. 276, 37093-37101
   Abstract »    Full Text »    PDF »
Effect of HIV integrase inhibitors on the RAG1/2 recombinase.
M. Melek, J. M. Jones, M. H. O'Dea, G. Pais, T. R. Burke Jr., Y. Pommier, N. Neamati, and M. Gellert (2002)
PNAS 99, 134-137
   Abstract »    Full Text »    PDF »
Intermediates in V(D)J recombination: A stable RAG1/2 complex sequesters cleaved RSS ends.
J. M. Jones and M. Gellert (2001)
PNAS 98, 12926-12931
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)