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Distinct and opposite diversifying activities of terminal transferase splice variants

Nature Immunology volume 3, pages 457–462 (2002)Cite this article

Abstract

The short splice variant of mouse terminal deoxynucleotidyl transferase (TdTS) catalyzes the addition of nontemplated nucleotides (N addition) at the coding joins of B cell and T cell antigen receptor genes. However, the activity and function of the long isoform of TdT (TdTL) have not been determined. We show here, in vitro and in vivo, that TdTL is a 3′→5′ exonuclease that catalyzes the deletion of nucleotides at coding joins. These findings suggest that the two TdT isoforms may act in concert to preserve the integrity of the variable region of antigen receptors while generating diversity.

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Figure 1: TdTL transcripts and protein are expressed in bone marrow pro-B and pre-B cells.
Figure 2: TdTL is a 3′→5′ exonuclease.
Figure 3: TdTS and TdTL modify a wide range of DNA substrates and alanine substitutions at carboxylate residues, markedly reduce the activity of TdTL.
Figure 4: TdTL deletes nucleotides from coding ends but not from signal ends in vivo.
Figure 5: Exonuclease core motifs are conserved in the amino acid sequences of murine and bovine TdTL.

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Acknowledgements

We thank S. Gilfillan and D. Mathis for the TdTL and TdTS cDNA clones; D. J. Mazure and F. W. Perrino for hTREX2; M. Oettinger for full-length RAG-1 and RAG-2; L. Gartland for FACS; X. Y. Liu for technical assistance; M. A. Anderson, D. S. Nelson, P. D. Burrows for helpful discussions; and A. Brookshire for preparing this manuscript. Supported by NIH grants AI 523133, AI 07051, AI36420 and the Howard Hughes Medical Institute (D. B. R.).

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Author notes

  1. Mary M. Purugganan

    Present address: Cain Project in Engineering and Professional Communication, Rice University, P.O. Box 1892, MS345, Houston, TX, 77251-1892, USA

Authors and Affiliations

  1. Division of Developmental and Clinical Immunology, Department of Microbiology, The University of Alabama at Birmingham, 378 Wallace Tumor Institute, Birmingham, 35294, AL, USA

    To-Ha Thai & John F. Kearney

  2. Department of Immunology, M929, Baylor College of Medicine, 1 Baylor Plaza, Houston, 77030, TX, USA

    Mary M. Purugganan & David B. Roth

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Correspondence to John F. Kearney.

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Thai, TH., Purugganan, M., Roth, D. et al. Distinct and opposite diversifying activities of terminal transferase splice variants. Nat Immunol 3, 457–462 (2002). https://doi.org/10.1038/ni788

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