Expression and V(D)J recombination activity of mutated RAG-1 proteins

Moshe J. Sadofsky, Joanne E. Hesse, J. Fraser McBlane, Martin Gellert

Research output: Contribution to journalArticle

165 Citations (Scopus)

Abstract

The products of the RAG-1 and RAG-2 genes ([1], [2]) are essential for the recombination of the DNA encoding the antigen receptors of the developing immune system. Little is known of the specific role these genes play. We have explored the sequences encoding mouse RAG-1 by deleting large parts of the gene and by introducing local sequence changes. We find that a RAG-1 gene with 40% of the coding region deleted still retains its recombination function. In addition, a series of small deletions within the strongly conserved remaining 60% of the coding region was tested. Nine out of ten of these prove unable to provide RAG-1 activity, but one is quite active. Certain peptide sequences were also specifically targeted for mutagenesis. The RAG-1 protein generated from this expression system is transported to the nucleus and is degraded with a 15 minute half-life. The fate of the proteins made by the deletion mutants were also assessed. Transport of RAG-1 protein to the nucleus was found even with the most extensive deletions studied. The functionality of the deleted proteins is discussed with relation to an alignment of RAG-1 sequences from five animal species.

Original languageEnglish (US)
Pages (from-to)5644-5650
Number of pages7
JournalNucleic Acids Research
Volume21
Issue number24
StatePublished - Dec 11 1993
Externally publishedYes

Fingerprint

RAG-1 Genes
V(D)J Recombination
Recombination
Genes
Deletion
Gene
Proteins
Protein
Genetic Recombination
Nucleus
Antigen Receptors
Encoding
Coding
Mutagenesis
Half-Life
Immune system
Immune System
Antigens
Mutant
Peptides

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Sadofsky, M. J., Hesse, J. E., McBlane, J. F., & Gellert, M. (1993). Expression and V(D)J recombination activity of mutated RAG-1 proteins. Nucleic Acids Research, 21(24), 5644-5650.

Expression and V(D)J recombination activity of mutated RAG-1 proteins. / Sadofsky, Moshe J.; Hesse, Joanne E.; McBlane, J. Fraser; Gellert, Martin.

In: Nucleic Acids Research, Vol. 21, No. 24, 11.12.1993, p. 5644-5650.

Research output: Contribution to journalArticle

Sadofsky, MJ, Hesse, JE, McBlane, JF & Gellert, M 1993, 'Expression and V(D)J recombination activity of mutated RAG-1 proteins', Nucleic Acids Research, vol. 21, no. 24, pp. 5644-5650.
Sadofsky MJ, Hesse JE, McBlane JF, Gellert M. Expression and V(D)J recombination activity of mutated RAG-1 proteins. Nucleic Acids Research. 1993 Dec 11;21(24):5644-5650.
Sadofsky, Moshe J. ; Hesse, Joanne E. ; McBlane, J. Fraser ; Gellert, Martin. / Expression and V(D)J recombination activity of mutated RAG-1 proteins. In: Nucleic Acids Research. 1993 ; Vol. 21, No. 24. pp. 5644-5650.
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