Ternary complex factors SAP-1 and Elk-1, but not net, are functionally equivalent in thymocyte development

J Immunol. 2010 Jul 15;185(2):1082-92. doi: 10.4049/jimmunol.1000472. Epub 2010 Jun 16.

Abstract

The ternary complex factors (TCFs; SAP-1, Elk-1, and Net) are serum response factor cofactors that share many functional properties and are coexpressed in many tissues. SAP-1, the predominant thymus TCF, is required for thymocyte positive selection. In this study, we assessed whether the different TCFs are functionally equivalent. Elk-1 deletion, but not the hypomorphic Net(delta) mutation, exacerbated the SAP-1 positive selection phenotype, but triply deficient thymocytes were no more defective than SAP-1(-/-) Elk-1(-/-) cells. Inactivation of the other TCFs did not affect SAP-1-independent processes, including beta-selection, regulatory T cell selection, and negative selection, although reduced marginal zone B cells were observed in SAP-1(-/-) Elk-1(-/-) animals. Ectopic expression of Elk-1, but not Net, rescued positive selection of SAP-1(-/-) thymocytes; thus, SAP-1 and Elk-1 are functionally equivalent in this system, and the SAP-1 null selection phenotype reflects only its high expression in the thymus. Array analysis of TCR-stimulated double-positive cells identified SAP-1-dependent inducible genes whose transcription was further impaired in SAP-1(-/-) Elk-1(-/-) cells; thus, these genes, which include Egr-1 and Egr-2, represent candidate mediators of positive selection. Chromatin immunoprecipitation revealed subtly different promoter targeting between the different TCFs. Ectopic expression of Egr-1 restored positive selection in SAP-1 null thymocytes, establishing it (and possibly other Egr family members) as the major effector for ERK-SAP-1 signaling in thymocyte positive selection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cell Differentiation / immunology
  • Cells, Cultured
  • Chromatin Immunoprecipitation
  • Female
  • Flow Cytometry
  • Gene Expression Profiling
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Oligonucleotide Array Sequence Analysis
  • Proto-Oncogene Proteins c-ets / genetics
  • Proto-Oncogene Proteins c-ets / immunology*
  • Proto-Oncogene Proteins c-ets / metabolism
  • Retroviridae / genetics
  • Thymus Gland / cytology
  • Thymus Gland / immunology*
  • Thymus Gland / metabolism
  • Transduction, Genetic
  • ets-Domain Protein Elk-1 / genetics
  • ets-Domain Protein Elk-1 / immunology*
  • ets-Domain Protein Elk-1 / metabolism
  • ets-Domain Protein Elk-4 / genetics
  • ets-Domain Protein Elk-4 / immunology*
  • ets-Domain Protein Elk-4 / metabolism

Substances

  • Elk1 protein, mouse
  • Elk3 protein, mouse
  • Proto-Oncogene Proteins c-ets
  • ets-Domain Protein Elk-1
  • ets-Domain Protein Elk-4

Associated data

  • GEO/GSE21546