Cachaco, S. | bioRxiv Science

bioRxiv2025

Manganese mediates antiviral effects by driving an ATM -TBK1 phosphorylation signaling pathway

We previously reported that manganese (Mn) enhances innate immune responses to viral infection by inducing phosphorylation of TANK-binding kinase 1 (TBK1) in an Ataxia-telangiectasia mutated (ATM)-dependent manner. However, the underlying mechanism by which how Mn induces TBK1 phosphorylation remained unclear. Here, we show that Mn dose-dependently induced TBK1 phosphorylation in the presence of ATM across multiple cell lines, as well as in primary human macrophages and T cells. This phosphorylation was abolished in ATM-deficient cells, and we identified cytoplasmic ATM as a key mediator. Immunoprecipitation assays revealed that Mn promoted ATM phosphorylation at Ser1893, Ser1981, and Ser2996. TBK1 interacted with phosphorylated ATM at early stages, but upon phosphorylation, TBK1 dissociated from the ATM-TBK1 complex. This dissociation coincided with enhanced antiviral cytokine production. Furthermore, Mn dose-dependently suppressed HIV replication by inducing multiple antiviral host factors and cytokines. Together, these findings identify a cytoplasmic ATM-TBK1 phosphorylation cycle as a critical regulator of antiviral innate immunity and suggest Mn supplementation as a potential therapeutic approach against HIV and other viral infections. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=111 SRC="FIGDIR/small/671272v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@1c72f62org.highwire.dtl.DTLVardef@1f7e789org.highwire.dtl.DTLVardef@820fe0org.highwire.dtl.DTLVardef@e06fb3_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical Abstract:C_FLOATNO Mn-dependent activation of the ATM-TBK1 phosphorylation signaling pathway. At early time points, Mn phosphorylated ATM at multiple site. And then TBK1 bound to p-ATM and became phosphorylated. Phosphorylated TBK1 then dissociated from the complex at later stages (right panel). P-TBK1 participated in activating downstream TBK1-IRF signaling, thereby enhancing antiviral cytokines induction to DNA or RNA virus infection (left panel). C_FIG

immunology↗

bioRxiv2024

Escherichia coli grown in cost-effective conical tubes produces more plasmid DNA

Before the COVID-19 pandemic, we grew plasmid-transformed Escherichia coli in Falcon round-bottom-polypropylene tubes (F-Round-PP) and isolated plasmid using a Miniprep kit. When obtaining sufficient quantities of F-Round-PP became problematic during the pandemic and the inflation, we grew them using any available tubes. Notably, we observed that plasmid yield from cells grown in a cost-effective Oxford conical-polypropylene tubes (O-Conical-PP) was higher than that from F-Round-PP. We assessed the impact using O-Conical-PP and other conical brand tubes. As a result, the plasmid yield from O-Conical-PP (the list price is nearly 1/3 of F-Round-PP) was 1.5-fold higher than other PP tubes (p<0.001). We propose that researchers may need to re-assess the effectiveness of their laboratory supplies to optimize the budget during this inflationary period.

microbiology↗