RAG-1 interacts with the repeated amino acid motif of the human homologue of the yeast protein SRP1.
Genes for immunoglobulins and T-cell receptor are generated by a process known as V(D)J recombination. This process is highly regulated and mediated by the recombination activating proteins RAG-1 and RAG-2. By the use of the two-hybrid protein interaction system, we isolated a human protein that specifically interacts with RAG-1. This ... protein is the human homologue of the yeast SRP1 (suppressor of a temperature-sensitive RNA polymerase I mutation). The SRP1-1 mutation is an allele-specific dominant suppressor of a temperature-sensitive mutation in the zinc binding domain of the 190-kDa subunit of Saccharomyces cerevisiae RNA polymerase I. The human SRP cDNA clone was used to screen a mouse cDNA library. We obtained a 3.9-kbp cDNA clone encoding the mouse SRP1. The open reading frame of this cDNA encodes a 538-amino acid protein with eight degenerate repeats of 40-45 amino acids each. The mouse and human SRP1 are 98% identical, while the mouse and yeast SRP1 have 48% identity. After cotransfection of the genes encoding RAG-1 and human SRP1 into 293T cells, a stable complex was evident. Deletion analysis indicated that the region of the SRP1 protein interacting with RAG-1 involved four repeats. The domain of RAG-1 that associates with SRP1 mapped N-terminal to the zinc finger domain. Because this region of RAG-1 is not required for recombination and SRP1 appears to be bound to the nuclear envelope, we suggest that this interaction helps to localize RAG-1.
Mesh Terms:
Amino Acid Sequence, Animals, DNA Mutational Analysis, Gene Rearrangement, T-Lymphocyte, Homeodomain Proteins, Humans, Mice, Molecular Sequence Data, Nuclear Proteins, Protein Binding, Proteins, Receptors, Antigen, T-Cell, Recombination, Genetic, Repetitive Sequences, Nucleic Acid, Saccharomyces cerevisiae, Sequence Homology, Amino Acid, Structure-Activity Relationship, alpha Karyopherins
Amino Acid Sequence, Animals, DNA Mutational Analysis, Gene Rearrangement, T-Lymphocyte, Homeodomain Proteins, Humans, Mice, Molecular Sequence Data, Nuclear Proteins, Protein Binding, Proteins, Receptors, Antigen, T-Cell, Recombination, Genetic, Repetitive Sequences, Nucleic Acid, Saccharomyces cerevisiae, Sequence Homology, Amino Acid, Structure-Activity Relationship, alpha Karyopherins
Proc. Natl. Acad. Sci. U.S.A.
Date: Aug. 02, 1994
PubMed ID: 8052633
View in: Pubmed Google Scholar
Download Curated Data For This Publication
7045
Switch View:
- Interactions 3