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Biology subjects

Miljkovic, J. L.

Publications and source records attributed to Miljkovic, J. L..

4 recordsLinked to original sources

TGS1 mediates mRNA 5'-cap trimethylation to promote oxidative phosphorylation in acute myeloid leukaemia.

The Trimethyl guanosine Synthase (TGS1) is a highly conserved enzyme mediating di-methylation of the 5-cap 7-methylguanosine of RNA to generate 2,2,7-trimethylguanosine (m2,2,7G). Known TGS1 targets include snRNAs, snoRNAs, the telomeric RNA component and a limited number of mRNAs encoding selenoproteins. TGS1 is highly expressed in acute myeloid leukaemia (AML) cells, and its expression correlates with poor prognosis. Here, we report that TGS1 directly methylates the cap of more than 500 mRNAs in AML cells. Specifically, we demonstrate that TGS1 modifies nuclear mRNAs encoding mitochondrial proteins, including critical components of complexes involved in both the TCA cycle and oxidative phosphorylation, promoting their translation mediated by mitochondria associated cytosolic ribosomes. Functionally, we report that TGS1 depletion impairs mitochondrial respiration and increases oxidative stress. This, in turn, impairs the growth of AML cells causing differentiation and cell cycle arrest in vitro and in vivo. Finally, we demonstrate that TGS1 depletion sensitizes AML cells to RSL3, a small molecule promoting ferroptosis. Taken together, our findings establish TGS1 as a key regulator of oxidative phosphorylation and mitochondrial redox status of AML cells and highlight its potential as a therapeutic target in leukaemia.

cancer biology↗

Protein thiol alterations drive aberrant phase separation in aging

Cellular homeostasis relies on precise regulation through chemical processes, such as protein posttranslational modifications (PTM) and physical processes, such as biomolecular condensation. Aging disrupts this balance, increasing susceptibility to diseases and death. However, the mechanisms behind age-related pathogenesis remain elusive. In this study, we dissected various cysteine PTMs and their impact on protein-mediated biomolecular condensation in aging brain. Our findings reveal that aging is associated with significant remodeling of cysteine PTMs, which impacts protein ability to participate in liquid-liquid phase separation (LLPS). Specifically, aging leads to an increase in protein sulfenylation and sulfonylation, which promotes LLPS and through conformational change increases the propensity of proteins to aggregate. Protein persulfidation, a protective thiol modification, prevents this by causing condensate dissolution. We demonstrate that age-induced alterations in cysteine PTMs influence the LLPS properties of synapsin-1 and G3BP2, resulting in disruptions in neurotransmitter release and stress granule formation, respectively. Additionally, our study uncovers that GAPDH is susceptible to LLPS and cysteine sulfonylation exacerbates its transition from condensates to aggregates. Mice deficient in cystathionine gamma-lyase, a pro-longevity gene that regulates intracellular persulfide levels, exhibit a shorter lifespan and spontaneous development of neurofibrillary tangles.

biochemistry↗

Naked mole rats have distinctive cardiometabolic and genetic adaptations to their underground low-oxygen lifestyles

The naked mole-rat Heterocephalus glaber is a eusocial mammal exhibiting extreme longevity (37-year lifespan), extraordinary resistance to hypoxia and absence of cardiovascular disease. To identify the mechanisms behind these exceptional traits, RNAseq and metabolomics of cardiac tissue from naked mole-rats was compared to other African mole-rat genera. We identified metabolic and genetic adaptations unique to naked mole-rats including elevated glycogen, thus enabling glycolytic ATP generation during cardiac ischemia. Elevated normoxic expression of HIF-1 was observed while downstream hypoxia responsive-genes were down regulated, suggesting adaptation to low oxygen environments. Naked mole-rat hearts showed reduced succinate build-up during ischemia and negligible tissue damage following ischemia-reperfusion injury. These adaptive evolutionary traits reflect a unique hypoxic and eusocial lifestyle that collectively may contribute to their longevity and health span. One Sentence SummaryNaked mole-rats have metabolic adaptations distinct from other subterranean genera rendering them resistant to cardiovascular pathology.

physiology↗

Disruption of the TCA cycle reveals an ATF4-dependent integration of redox and amino acid metabolism

The Tricarboxylic Acid Cycle (TCA) cycle is arguably the most critical metabolic cycle in physiology and exists as an essential interface coordinating cellular metabolism, bioenergetics, and redox homeostasis. Despite decades of research, a comprehensive investigation into the consequences of TCA cycle dysfunction remains elusive. Here, we targeted two TCA cycle enzymes, fumarate hydratase (FH) and succinate dehydrogenase (SDH), and combined metabolomics, transcriptomics, and proteomics analyses to fully appraise the consequences of TCA cycle inhibition (TCAi) in kidney epithelial cells. Our comparative approach shows that TCAi elicits a convergent rewiring of redox and amino acid metabolism dependent on the activation of ATF4 and the integrated stress response (ISR). Furthermore, we also uncover a divergent metabolic response, whereby acute FHi, but not SDHi, can maintain asparagine levels via reductive carboxylation and maintenance of cytosolic aspartate synthesis. Our work highlights an important interplay between the TCA cycle, redox biology and amino acid homeostasis. HighlightsO_LITCA cycle inhibition promotes GSH synthesis and impairs de novo aspartate and proline synthesis C_LIO_LIDisruption of mitochondrial thiol redox homeostasis phenocopies TCA cycle inhibition by promoting GSH synthesis and impairing proline and aspartate synthesis C_LIO_LIAcute FHi, but not SDHi, can maintain asparagine levels via reductive carboxylation and maintenance of cytosolic aspartate synthesis C_LIO_LITCA cycle inhibition mimics an amino acid deprivation-type response and activates ATF4 via the integrated stress response to maintain redox and amino acid homeostasis C_LI

biochemistry↗