rag-1 and rag-2 are components of a high-molecular-weight complex, and association of rag-2 with this complex is rag-1 dependent.
Despite the essential and synergistic functions of the rag-1 and rag-2 proteins in V(D)J recombination and lymphocyte development, little is known about the biochemical properties of the two proteins. We have developed cell lines expressing high levels of the rag proteins and specific, sensitive immunological reagents for their detection, and we have examined the physical properties of the rag proteins in vitro and their subcellular localizations in vivo. rag-1 is tightly associated with nuclear structures, requires a high salt concentration to maintain its solubility, and is a component of large, heterogeneously sized complexes. Furthermore, the presence of rag-1 alters the behavior of rag-2, conferring on it properties similar to those of rag-1 and changing its distribution in the nucleus. We demonstrate that rag-1 and rag-2 are present in the same complex by coimmunoprecipitation, and we provide evidence that these complexes contain more molecules of rag-2 than of rag-1. The demonstration of intracellular complexes containing rag-1 and rag-2 raises the possibility that interaction between these proteins is necessary for their biological function.
Leu
T M
TM
Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
Schatz
D G
DG
D000818:Animals; D001483:Base Sequence; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000737:chemistry; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D016393:Lymphoma, B-Cell; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D011995:Recombination, Genetic / Q000502:physiology*; D013950:Thymus Gland / Q000737:chemistry; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D011506:Proteins; C499308:Rag2 protein, mouse; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein