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Control of thymocyte development and recombination-activating gene expression by the zinc finger protein Zfp608

Nature Immunology volume 7, pages 1309–1316 (2006)Cite this article

Abstract

The products of recombination-activating gene 1 (Rag1) and Rag2 are required for T cell receptor gene assembly and thymocyte maturation, yet their transcriptional control mechanisms remain unclear. A congenic strain (called 'ZORI' here) with defects in Rag1 and Rag2 expression, thymocyte maturation and peripheral T cell homeostasis has been developed. Here, we mapped the mutation in this strain to a chromosome 18 locus containing a single known gene encoding the zinc finger protein Zfp608. This gene (Zfp608) was highly expressed in neonatal thymus but was extinguished thereafter. In contrast to wild-type mice, ZORI mice had sustained thymocyte expression of Zfp608 throughout life. The ZORI mutation produced a thymocyte-intrinsic developmental defect. Overexpression of Zfp608 in BALB/c thymocytes substantially impaired Rag1 and Rag2 expression, indicating the underlying mechanism for the defect in ZORI thymocyte development. Thus, the normal function of Zfp608 may be to prevent Rag1 and Rag2 expression in utero.

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Figure 1: Wasting syndrome, premature mortality and T cell immunodeficiency in homozygous ZORI mice.
Figure 2: Thymocyte subpopulations in BALB/c and ZORI mice.
Figure 3: Overexpression of Zfp608 mRNA in ZORI thymus.
Figure 4: Expression of Notch1, Hes1, Jag1 and Jag2 by ZORI and BALB/c mice.
Figure 5: Reconstitution of bone marrow cells in lethally irradiated mice.
Figure 6: Zfp608 negatively regulates TCR recombination and Rag1 and Rag2 expression.
Figure 7: Thymic expression of Zfp608 mRNA is inversely correlated with thymocyte numbers during adolescence.

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Acknowledgements

We thank J.W. Thomas and L. Van Kaer for comments and review of the manuscript, and J. Sligh for comments and assistance with the temperature-gradient capillary electrophoresis technique; M. Boothby (Vanderbilt University) provided OTII OVA-specific TCR-transgenic mice. Supported by the US National Institutes of Health (5 T32 DK07563 to F.Z., AI 44924 to T.M.A., P01 HL68744 and CA100905 to E.M.O., and F32 AI066691 and 5T32 HL069765 to L.R.T.).

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Authors and Affiliations

  1. Division of Rheumatology, Department of Medicine, Vanderbilt University School of Medicine, Nashville, 37232, Tennessee, USA

    Feng Zhang & Thomas M Aune

  2. Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, 37232, Tennessee, USA

    Feng Zhang, Lance R Thomas, Eugene M Oltz & Thomas M Aune

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  1. Feng Zhang
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Contributions

All authors contributed to discussions, experimental design, data analysis and preparation of the manuscript; F.Z. designed and executed experiments and prepared the manuscript; L.R.T. examined TCR rearrangement and helped to analyze data and to write the manuscript; and T.M.A. and E.M.O. supervised the work and assisted in data analysis and composition of the manuscript.

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Correspondence to Thomas M Aune.

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Supplementary information

Supplementary Fig. 1

T cell development defects in the ZORI stain map to a locus between 54.3 Mb (D18mit150) and 55.8 Mb (D18mit218) on chromosome 18. (PDF 307 kb)

Supplementary Fig. 2

Relative expression of genes adjacent to Zfp608 on chromosome 18. (PDF 260 kb)

Supplementary Note 1

MapPair primer sequences and primer sequences for TCR rearrangements. (PDF 7 kb)

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Zhang, F., Thomas, L., Oltz, E. et al. Control of thymocyte development and recombination-activating gene expression by the zinc finger protein Zfp608. Nat Immunol 7, 1309–1316 (2006). https://doi.org/10.1038/ni1397

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