Leaky severe combined immunodeficiency in mice lacking non-homologous end joining factors XLF and MRI.

Sergio Castañeda-Zegarra,Marion Fernandez-Berrocal,Valentyn Oksenych,Qindong Zhang,Rouan Yao,Amin Alirezaylavasani

doi:10.18632/aging.202346

Abstract

Non-homologous end-joining (NHEJ) is a DNA repair pathway required to detect, process, and ligate DNA double-stranded breaks (DSBs) throughout the cell cycle. The NHEJ pathway is necessary for V(D)J recombination in developing B and T lymphocytes. During NHEJ, Ku70 and Ku80 form a heterodimer that recognizes DSBs and promotes recruitment and function of downstream factors PAXX, MRI, DNA-PKcs, Artemis, XLF, XRCC4, and LIG4. Mutations in several known NHEJ genes result in severe combined immunodeficiency (SCID). Inactivation of Mri, Paxx or Xlf in mice results in normal or mild phenotype, while combined inactivation of Xlf/Mri, Xlf/Paxx, or Xlf/Dna-pkcs leads to late embryonic lethality. Here, we describe three new mouse models. We demonstrate that deletion of Trp53 rescues embryonic lethality in mice with combined deficiencies of Xlf and Mri. Furthermore, Xlf-/-Mri-/-Trp53+/- and Xlf-/-Paxx-/-Trp53+/- mice possess reduced body weight, severely reduced mature lymphocyte counts, and accumulation of progenitor B cells. We also report that combined inactivation of Mri/Paxx results in live-born mice with modest phenotype, and combined inactivation of Mri/Dna-pkcs results in embryonic lethality. Therefore, we conclude that XLF is functionally redundant with MRI and PAXX during lymphocyte development in vivo. Moreover, Mri genetically interacts with Dna-pkcs and Paxx.

Highlights

Non-homologous end-joining (NHEJ) is a DNA repair pathway that recognizes, processes and ligates DNA double-stranded breaks (DSB) throughout the cell cycle
Recent findings by our and other research groups suggest that MRI forms heterogeneous complexes involving paralogue of XRCC4 and XLF (PAXX) or XRCC4-like factor (XLF), which function during DNA DSB repair by NHEJ [5]
Combined inactivation of Xlf and Mri [5] or Xlf and Paxx [4, 14, 15] results in embryonic lethality, which correlates with increased levels of neuronal apoptosis in the central nervous system (CNS) (Figure 5)

Summary

Introduction

Non-homologous end-joining (NHEJ) is a DNA repair pathway that recognizes, processes and ligates DNA double-stranded breaks (DSB) throughout the cell cycle. NHEJ is required for lymphocyte development; in particular, to repair DSBs induced by the recombination activating genes (RAG) 1 and 2 in developing B and T lymphocytes, and by activation-induced cytidine deaminase (AID) in mature B cells [1]. NHEJ is initiated when Ku70 and Ku80 (Ku) are recruited to the DSB sites. Together with DNA-dependent protein kinase, catalytic subunit (DNA-PKcs), forms the DNAPK holoenzyme [2]. The nuclease Artemis is recruited to the DSB sites to process DNA hairpins and overhangs [3].

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Results

Conclusion