Isolation of interleukin 2-induced immediate-early genes

Carol Beadling, Kirk W. Johnson, Kendall A. Smith

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

Clonal expansion of antigen-reactive T lymphocytes is driven by the lymphokine interleukin 2 (IL-2). To further elucidate the mechanisms of IL-2 action, we have utilized a differential hybridization procedure to clone IL-2-induced immediate-early genes from an IL-2-stimulated human T-cell cDNA library. To increase the frequency of IL-2-induced transcripts represented in the library, the protein synthesis inhibitor cycloheximide was included during the 2-hr IL-2 stimulation to superinduce gene expression, and the uridine analogue 4-thiouridine was utilized to enable selective purification of newly synthesized transcripts. From the enriched library, we have isolated eight IL-2-induced genes, six of which represent previously unrecognized human sequences. Northern blot analysis revealed that the induction of seven of the genes is specific to the IL-2-mediated G1 "progression" phase of the cell cycle, in that only one gene is also induced during the T-cell receptor-triggered G0-G1 "competence" phase. These results indicate that the effects of IL-2 are mediated by the specific induction of a number of immediate-early genes and provide a means with which to further delineate the mechanisms whereby IL-2 stimulates T-lymphocyte proliferation and differentiation. The methods described in this report should also be of general utility in the dissection of the signaling pathways activated by diverse cytokine receptors.

Original languageEnglish (US)
Pages (from-to)2719-2723
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number7
StatePublished - Apr 1 1993
Externally publishedYes

Fingerprint

Immediate-Early Genes
Interleukin-2
G1 Phase
T-Lymphocytes
Libraries
Thiouridine
Genes
Cytokine Receptors
Protein Synthesis Inhibitors
Lymphokines
Uridine
Cycloheximide
T-Cell Antigen Receptor
Gene Library
Northern Blotting
Mental Competency
Dissection
Cell Cycle
Clone Cells
Gene Expression

Keywords

  • Cytokine-responsive genes
  • Differential cloning
  • Human T cells

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Isolation of interleukin 2-induced immediate-early genes. / Beadling, Carol; Johnson, Kirk W.; Smith, Kendall A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 7, 01.04.1993, p. 2719-2723.

Research output: Contribution to journalArticle

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