Notch-induced endoplasmic reticulum-associated degradation governs mouse thymocyte β- selection
- Baylor College of Medicine, United States
- Case Western Reserve University, United States
- Wayne State University, United States
- La Trobe University, Australia
- University of Michigan-Ann Arbor, United States
Abstract
Signals from the pre-T cell receptor and Notch coordinately instruct b-selection of CD4-CD8- double negative (DN) thymocytes to generate ab T cells in the thymus. However, how these signals ensure a high-fidelity proteome and safeguard the clonal diversification of the pre-selection TCR repertoire given the considerable translational activity imposed by b-selection is largely unknown. Here, we identify the endoplasmic reticulum (ER)-associated degradation (ERAD) machinery as a critical proteostasis checkpoint during b-selection. Expression of the SEL1L-HRD1 complex, the most conserved branch of ERAD, is directly regulated by the transcriptional activity of the Notch intracellular domain. Deletion of Sel1l impaired DN3 to DN4 thymocyte transition and severely impaired mouse ab T cell development. Mechanistically, Sel1l deficiency induced unresolved ER stress that triggered thymocyte apoptosis through the PERK pathway. Accordingly, genetically inactivating PERK rescued T cell development from Sel1l-deficient thymocytes. In contrast, IRE1a/XBP1 pathway was induced as a compensatory adaptation to alleviate Sel1l-deficiency induced ER stress. Dual loss of Sel1l and Xbp1 markedly exacerbated the thymic defect. Our study reveals a critical developmental signal controlled proteostasis mechanism that enforces T cell development to ensure a healthy adaptive immunity.
Data availability
Sequencing data have been deposited in GEO under accession code GSE173993.All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all Figures.
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Notch-Induced Endoplasmic Reticulum-Associated Degradation Governs Thymocyte Beta-SelectionNCBI Gene Expression Omnibus, GSE173993.
Article and author information
Author details
Katharine Umphred-Wilson
Funding
National Heart, Lung, and Blood Institute (R01HL146642)
- Xi Chen
Cancer Prevention and Research Institute of Texas (RP160283)
- Fanglue Peng
National Institute of Allergy and Infectious Diseases (R01 AI1143992)
- Stanley Adoro
National Cancer Institute (R37CA228304)
- Xi Chen
National Cancer Institute (K22CA218467)
- Stanley Adoro
National Cancer Institute (P50CA186784)
- Xi Chen
National Institute of General Medical Sciences (R35GM130292)
- Ling Qi
DOD Peer Reviewed Cancer Research Program (W81XWH1910524)
- Xi Chen
DOD Peer Reviewed Cancer Research Program (W81XWH1910306)
- Stanley Adoro
Congressionally Directed Medical Research Programs (W81XWH1910035)
- Xiangdong Lv
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Juan Carlos Zúñiga-Pflücker, University of Toronto, Sunnybrook Research Institute, Canada
Ethics
Animal experimentation: All protocols described in this study were approved by the Baylor College of Medicine Institutional Animal Care and Use Committee (protocol: AN-6813) or Case Western Reserve University Institutional Animal Care and Use Committee (protocol: 2017-0055).
Version history
- Received: May 3, 2021
- Preprint posted: May 7, 2021 (view preprint)
- Accepted: July 5, 2021
- Accepted Manuscript published: July 9, 2021 (version 1)
- Version of Record published: July 27, 2021 (version 2)
- Version of Record updated: July 28, 2021 (version 3)
Copyright
© 2021, Liu et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Notch-induced endoplasmic reticulum-associated degradation governs mouse thymocyte β- selection
eLife 10:e69975.
https://doi.org/10.7554/eLife.69975
Further reading
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- Chromosomes and Gene Expression
- Developmental Biology
Developmental programming involves the accurate conversion of signalling levels and dynamics to transcriptional outputs. The transcriptional relay in the Notch pathway relies on nuclear complexes containing the co-activator Mastermind (Mam). By tracking these complexes in real time, we reveal that they promote the formation of a dynamic transcription hub in Notch ON nuclei which concentrates key factors including the Mediator CDK module. The composition of the hub is labile and persists after Notch withdrawal conferring a memory that enables rapid reformation. Surprisingly, only a third of Notch ON hubs progress to a state with nascent transcription, which correlates with polymerase II and core Mediator recruitment. This probability is increased by a second signal. The discovery that target-gene transcription is probabilistic has far-reaching implications because it implies that stochastic differences in Notch pathway output can arise downstream of receptor activation.
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- Developmental Biology
Large-scale cell flow characterizes gastrulation in animal development. In amniote gastrulation, particularly in avian gastrula, a bilateral vortex-like counter-rotating cell flow, called ‘polonaise movements’, appears along the midline. Here, through experimental manipulations, we addressed relationships between the polonaise movements and morphogenesis of the primitive streak, the earliest midline structure in amniotes. Suppression of the Wnt/planar cell polarity (PCP) signaling pathway maintains the polonaise movements along a deformed primitive streak. Mitotic arrest leads to diminished extension and development of the primitive streak and maintains the early phase of the polonaise movements. Ectopically induced Vg1, an axis-inducing morphogen, generates the polonaise movements, aligned to the induced midline, but disturbs the stereotypical cell flow pattern at the authentic midline. Despite the altered cell flow, induction and extension of the primitive streak are preserved along both authentic and induced midlines. Finally, we show that ectopic axis-inducing morphogen, Vg1, is capable of initiating the polonaise movements without concomitant PS extension under mitotic arrest conditions. These results are consistent with a model wherein primitive streak morphogenesis is required for the maintenance of the polonaise movements, but the polonaise movements are not necessarily responsible for primitive streak morphogenesis. Our data describe a previously undefined relationship between the large-scale cell flow and midline morphogenesis in gastrulation.
