bioRxiv Science⌕ Search

bioRxiv · 10.1101/2022.09.04.506508

Pseudokinase HPO-11 inhibits nonsense-mediated decay to ensure genome stability in C. elegans

Abstract

DNA double-strand breaks (DSBs) are highly toxic DNA lesions that can induce mutations and chromosome rearrangement therefore causing genome instability (GIN). In response to DSBs, cells activate the DNA damage response by hierarchical assembly of signaling and repair mechanisms. This involves recruitment of the repair factors at DSB sites, local chromatin remodeling, cell cycle arrest and, eventually, DNA repair or apoptosis. Studies investigating mechanisms ensuring genome stability have so far mostly focused on DNA-protein interactions and signal transduction in response to DNA damage. Emerging evidence in the last decade suggests that post-transcriptional control of gene expression by RNA-binding proteins also participates in maintaining genome integrity. However, how specific control of RNA fate mechanistically affects genome stability is still poorly understood. Here, we report that the pseudokinase HPO-11 ensures genome integrity in C. elegans. Loss of hpo-11 leads to accumulation of R-loops, increased DSBs and germline apoptosis, as well as an elevated mutation rate in the somatic cells. In addition, inhibition of nonsense mediated decay (NMD) reduces DSBs and germline apoptosis in the absence of hpo-11. We find that HPO-11 physically interacts with SMG-2, the core factor of NMD, and prevents degradation of specific transcripts by NMD, thus contributing to maintenance of genome stability. Furthermore, knock-down of hpo-11 human homologs NRBP1/2 also results in increased DNA DSBs, and NRBP1/2 physically interact with the human SMG-2 orthologue UPF1. In summary, our work identifies an evolutionarily conserved role of HPO-11 to protect genome stability via suppressing abnormal mRNA decay by NMD.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Zhao, Q., Gromoff, E. D. v., Yang, W., Schwarz, J., Schulze, E., Warscheid, B., Baumeister, R., Qi, W.. 2022-09-04. Pseudokinase HPO-11 inhibits nonsense-mediated decay to ensure genome stability in C. elegans. https://doi.org/10.1101/2022.09.04.506508

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Hypoxic Stress Induces LARP1-Driven Stress Granules That Protect mRNAs from Decay to Sustain Mitotic Cell Cycle Progression

Hypoxia is a defining feature of solid tumours that elicits extensive transcriptional and post-transcriptional adaptations. Stress granules (SGs) are frequently observed under hypoxia, yet how they assemble and whether they contribute to hypoxic gene regulation remain poorly understood. Here, we show that hypoxia induces SGs through an assembly mechanism distinct from that of classical acute stresses. Hypoxia SG formation occurs independently of the integrated stress response and instead requires the HIF1-REDD1-mTORCC1-4EBP1 axis. Remarkably, hypoxia SGs assemble despite robust ongoing translation, suggesting that additional mechanisms lower the threshold for RNA condensation. We identify LARP1 as a potential determinant of this process, where it promotes G3BP-dependent SG assembly specifically during hypoxia. Despite their distinct assembly mechanism, hypoxia and arsenite SGs recruit broadly similar RNA populations, with selective differences validated by single-molecule FISH, suggesting that conserved principles govern RNA partitioning across different SG states. Functionally, sequestration into hypoxia SGs correlates with enhance protein expression whereby a subset of mRNAs are protected from decay. Transcriptomic analyses further reveal that this protection sustains a broader gene-expression program enriched for MCM complex and mitotic cell-cycle regulators. Accordingly, disruption of hypoxia SGs reduces mitotic progression and proliferative competence during prolonged hypoxia. Together, our findings establish LARP1-mediated condensation as a mechanism that enables SG assembly despite ongoing translation and reveal that SG sequestration preserves selected mRNAs from decay to sustain the gene-expression programs required for long-term adaptation to hypoxia.

molecular biology↗

Age-associated repression of HNF4α is a driver and potential therapeutic target in liver metabolic dysfunction

Aging is a major risk factors for metabolic dysfunction-associated steatotic liver disease (MASLD). However, the mechanisms by which aging disrupts hepatocyte transcriptional networks to drive metabolic dysfunction remain poorly understood. Hepatocyte nuclear factor 4 alpha (HNF4) is a master regulator of hepatocyte identity and metabolic homeostasis, but its role in liver aging has not been defined. Here, we perform a comprehensive multi-omic profiling of young and aged mouse livers combined with targeted HNF4 functional studies to define how aging remodels HNF4-dependent gene regulation and whether HNF4 can be a target to reverse age-associated liver dysfunction. Despite preserved HNF4 expression with age, aging was associated with reduced HNF4 chromatin occupancy and transcriptional activity, accompanied by epigenetic remodeling, including changes in histone acetylation, chromatin accessibility, and DNA methylation. Integration and comparison of HNF4 CUT&RUN with hepatocyte-specific Hnf4a deletion in young and aged livers identified high-confidence overlapping but distinct young- and old- specific HNF4 target gene networks. In young hepatocytes, HNF4 coordinates two complementary transcriptional programs: activation of genes governing hepatocyte identity, lipid metabolism, and metabolic function, while simultaneously inhibiting alternative developmental, proliferative, and inflammatory programs. Spatial transcriptomic analyses further revealed zone-specific specialization of HNF4-dependent gene programs. Aging selectively disrupted these regulatory networks, resulting in compromised liver identity and metabolic gene programs, de-repression of inflammatory pathways and alternative cellular identity programs and disrupted hepatocyte zonation, consistent with age-dependent lipidomic and metabolic changes, hepatic fat accumulation, and immune infiltration. Hepatocyte-specific Hnf4a deletion in young mice recapitulated key features of liver aging, including impaired hepatocyte identity, metabolic dysfunction, steatosis, inflammatory activation and altered hepatic immune-cell composition, while deletion in aged mice further exacerbated steatosis, inflammation, and hepatic dysfunction. Conversely, hepatocyte-specific HNF4 restoration in aged mice significantly reduced hepatic steatosis and lipid accumulation, restored fatty acid metabolic programs, and partially reversed age-associated transcriptional changes. However, inflammatory and interferon-associated programs were not uniformly reversed, revealing differential responsiveness of metabolic and inflammatory pathways to HNF4 restoration. Together, these findings identify age-associated impairment of HNF4 activity as a functional contributor to liver aging, linking dysregulation of hepatocyte identity, metabolic homeostasis, and inflammatory control to age-associated hepatic dysfunction. Importantly, the selective reversibility of metabolic defects following HNF4 restoration establishes HNF4 as a promising strategy for preserving hepatocyte homeostasis and reversing age-associated hepatic metabolic dysfunction.

molecular biology↗

A Human Tumor Virus Recruits HSP70 Chaperone to Remodel Its Replisome on DNA

Human Merkel cell polyomavirus (MCV) large T (LT) protein functions as a replication helicase analogous to the cellular MCM complex but requires a second MCV protein, small T (sT), for efficient LT-initiated replication. Both LT and sT encode highly-conserved DnaJ domains that bind to the cellular refolding molecule, heat shock protein 70 (HSP70). Using single-molecule imaging and FRET, we find that sT recruits dimeric HSP70 to serve as a molecular bridge between sT and LT on DNA and that this complex facilitates remodeling LT into a functional double-hexameric helicase. Disruption of LT-HSP70-HSP70-sT interaction with HSP70 inhibitors or by mutations to the LT-sT DnaJ domains collapse the LT-HSP70-HSP70-sT complex, abolishing sT-enhanced LT dodecamerization and ATP-dependent DNA unwinding. Assembly of virus-free mammalian MCM3, MCM4 and MCM6 in nuclear extracts similarly recruits HSP70 to DNA suggesting that Hsp70 remodeling is used for efficient eukaryotic assembly of the multi-protein replication machinery.

molecular biology↗