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Biology subjects

Seibert, M.

Publications and source records attributed to Seibert, M..

3 recordsLinked to original sources

Extracellular protein catabolism drives regulated nitrogen handling and ammonia buffering in acute myeloid leukemia

Acute myeloid leukemia (AML) cells exhibit pronounced metabolic plasticity, yet how amino acid supply is coordinated to sustain leukemic metabolism remains poorly understood. Here, we show that AML cells catabolize extracellular proteins as a major source of amino acids through lysosomal degradation of albumin. This proteocatabolic activity supports anabolic processes and mitochondrial energy production and establishes a regulated, high-throughput regime of primary nitrogen-containing metabolites (nitrogen regimen). Sustained proteocatabolism inevitably generates ammonia, and we find elevated ammonia concentrations in bone marrow plasma from newly diagnosed AML patients that decline with effective induction therapy. Using metabolomics, isotope tracing and targeted genetic and pharmacological manipulations, we identify glutamate-ammonia ligase (GS/GLUL) as a central enzyme that buffers proteocatabolism-derived ammonia by stabilizing intracellular nitrogen homeostasis. Loss of GS function limits sustainable nitrogen handling capacity, thereby impairing leukemic proliferation and delaying disease progression in vivo. Together, our findings define extracellular protein catabolism as a regulating nitrogen management strategy in AML and reveal GS as a capacity-defining vulnerability of proteocatabolic growth.

cancer biology↗

Isoform-specific regulation of SMARCAD1 dosage in ectodermal homeostasis and dysplasia

Haploinsufficiency is a common cause of developmental disorders, yet the mechanisms regulating protein abundance of dosage-sensitive genes remain incompletely understood. SMARCAD syndrome comprises inherited ectodermal dysplasias caused by mutations affecting SMARCAD1-s, the skin-specific short isoform of the ATP-dependent chromatin remodeller SMARCAD1. Here, we show that independent patient-derived mutations impair splicing of this isoform, resulting in intron retention. Although total mRNA abundance is reduced, spliced transcripts remain translationally competent and yield variably reduced levels of wild-type protein, providing evidence for a threshold-dependent haploinsufficiency model of disease. Under physiological conditions, the isoform-specific non-coding exon 1 dampens protein production without altering transcript levels, revealing previously unrecognized post-transcriptional control of SMARCAD1-s dosage. Our findings implicate upstream open reading frames as candidate mediators of translational control. Unexpectedly, SMARCAD1-s exhibits regulated nucleocytoplasmic distribution. Functional analyses identify two nuclear localization signals that differentially contribute to localization of the SMARCAD1 isoforms. Together, our findings clarify the molecular basis of SMARCAD syndrome and identify post-transcriptional and spatial mechanisms controlling the abundance and localization of a dosage-sensitive chromatin remodeller, providing insight into protein dosage control.

molecular biology↗

Electrospray sample injection for single-particle imaging with X-ray lasers

The possibility of imaging single proteins constitutes an exciting challenge for X-ray lasers. Despite encouraging results on large particles, imaging small particles has proven to be difficult for two reasons: not quite high enough pulse intensity from currently available X-ray lasers and, as we demonstrate here, contamination of the aerosolised molecules by non-volatile contaminants in the solution. The amount of contamination on the sample depends on the initial droplet-size during aerosolisation. Here we show that with our electrospray injector we can decrease the size of aerosol droplets and demonstrate virtually contaminant-free sample delivery of organelles, small virions, and proteins. The results presented here, together with the increased performance of next generation X-ray lasers, constitute an important stepping stone towards the ultimate goal of protein structure determination from imaging at room temperature and high temporal resolution.

biophysics↗