This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Bailin, T.
Right arrow Articles by Sadofsky, M. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bailin, T.
Right arrow Articles by Sadofsky, M. J.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, July 1999, p. 4664-4671, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A RAG1 and RAG2 Tetramer Complex Is Active in Cleavage in V(D)J Recombination

Tu Bailin, Xianming Mo, and Moshe J. Sadofsky*

Program of Gene Regulation, Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, Georgia 30912-2650

Received 29 January 1999/Returned for modification 22 February 1999/Accepted 1 April 1999

During V(D)J recombination two proteins, RAG1 and RAG2, assemble as a protein-DNA complex with the appropriate DNA targets containing recombination signal sequences (RSSs). The properties of this complex require a fairly elaborate set of protein-protein and protein-DNA contacts. Here we show that a purified derivative of RAG1, without DNA, exists predominantly as a homodimer. A RAG2 derivative alone has monomer, dimer, and larger forms. The coexpressed RAG1 and RAG2 proteins form a mixed tetramer in solution which contains two molecules of each protein. The same tetramer of RAG1 and RAG2 plus one DNA molecule is the form active in cleavage. Additionally, we show that both DNA products following cleavage can still be held together in a stable protein-DNA complex.

* Corresponding author. Mailing address: Medical College of Georgia, Institute of Molecular Medicine and Genetics, Program of Gene Regulation, CB-2803, Augusta, GA 30912-2650. Phone: (706) 721-8761. Fax: (706) 721-8752. E-mail: moshe{at}immag.mcg.edu.

Molecular and Cellular Biology, July 1999, p. 4664-4671, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Giblin, W., Chatterji, M., Westfield, G., Masud, T., Theisen, B., Cheng, H.-L., DeVido, J., Alt, F. W., Ferguson, D. O., Schatz, D. G., Sekiguchi, J. (2009). Leaky severe combined immunodeficiency and aberrant DNA rearrangements due to a hypomorphic RAG1 mutation. Blood 113: 2965-2975 [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]  
  • 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]  
  • Swanson, P. C., Volkmer, D., Wang, L. (2004). Full-length RAG-2, and Not Full-length RAG-1, Specifically Suppresses RAG-mediated Transposition but Not Hybrid Joint Formation or Disintegration. J. Biol. Chem. 279: 4034-4044 [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]  
  • 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]  
  • 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]  
  • Huye, L. E., Purugganan, M. M., Jiang, M.-M., Roth, D. B. (2002). Mutational Analysis of All Conserved Basic Amino Acids in RAG-1 Reveals Catalytic, Step Arrest, and Joining-Deficient Mutants in the V(D)J Recombinase. Mol. Cell. Biol. 22: 3460-3473 [Abstract] [Full Text]  
  • Mundy, C. L., Patenge, N., Matthews, A. G. W., Oettinger, M. A. (2002). Assembly of the RAG1/RAG2 Synaptic Complex. Mol. Cell. Biol. 22: 69-77 [Abstract] [Full Text]  
  • Landree, M. A., Kale, S. B., Roth, D. B. (2001). Functional Organization of Single and Paired V(D)J Cleavage Complexes. Mol. Cell. Biol. 21: 4256-4264 [Abstract] [Full Text]  
  • Sadofsky, M. J. (2001). The RAG proteins in V(D)J recombination: more than just a nuclease. Nucleic Acids Res 29: 1399-1409 [Abstract] [Full Text]  
  • Mo, X., Bailin, T., Sadofsky, M. J. (2001). A C-Terminal Region of RAG1 Contacts the Coding DNA during V(D)J Recombination. Mol. Cell. Biol. 21: 2038-2047 [Abstract] [Full Text]  
  • Swanson, P. C. (2001). 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. Mol. Cell. Biol. 21: 449-458 [Abstract] [Full Text]  
  • Yu, K., Lieber, M. R. (2000). The Nicking Step in V(D)J Recombination Is Independent of Synapsis: Implications for the Immune Repertoire. Mol. Cell. Biol. 20: 7914-7921 [Abstract] [Full Text]  
  • Lew, S., Franco, D., Chang, Y. (2000). Activation of V(D)J Recombination Induces the Formation of Interlocus Joints and Hybrid Joints in scid Pre-B-Cell Lines. Mol. Cell. Biol. 20: 7170-7177 [Abstract] [Full Text]  
  • Agard, E. A., Lewis, S. M. (2000). Postcleavage Sequence Specificity in V(D)J Recombination. Mol. Cell. Biol. 20: 5032-5040 [Abstract] [Full Text]  
  • Brown, M. L., Chang, Y. (2000). Metabolism of Recombination Coding Ends in scid Cells. J. Immunol. 164: 4135-4142 [Abstract] [Full Text]  
  • Mo, X., Bailin, T., Noggle, S., Sadofsky, M. J. (2000). A highly ordered structure in V(D)J recombination cleavage complexes is facilitated by HMG1. Nucleic Acids Res 28: 1228-1236 [Abstract] [Full Text]  
  • Landree, M. A., Wibbenmeyer, J. A., Roth, D. B. (1999). Mutational analysis of RAG1 and RAG2 identifies three catalytic amino acids in RAG1 critical for both cleavage steps of V(D)J recombination. Genes Dev. 13: 3059-3069 [Abstract] [Full Text]  
  • Binnie, A., Olson, S., Wu, G. E., Lewis, S. M. (1999). Gamma-Irradiation Directly Affects the Formation of Coding Joints in SCID Cell Lines. J. Immunol. 163: 5418-5426 [Abstract] [Full Text]  
  • Lee, K.-J., Huang, J., Takeda, Y., Dynan, W. S. (2000). DNA Ligase IV and XRCC4 Form a Stable Mixed Tetramer That Functions Synergistically with Other Repair Factors in a Cell-free End-joining System. J. Biol. Chem. 275: 34787-34796 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»