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It is constitutively active at a low level in eukaryotic cells and can be induced by conditions of cellular stress, such as nutrient starvation. Moreover, autophagy plays an important role in diverse processes such as immunobiology, pathogen infection, ageing, and neurodegenerative and other diseases. Using a high\u2010content fluorescence assay for microtubule\u2010associated protein 1 light chain 3 beta (LC3B), a major player in the autophagic pathway, we screened a library of commercial drugs and natural products for activators and inhibitors of LC3B\u2010positive vesicle accumulation. Positive hits for known autophagy modulators included anisomycin, amphotericin B, carbonyl cyanide m\u2010chlorophenylhydrazone (CCCP) and cytochalasin D. Importantly, we identified several new autophagy modulators, such as aciclovir and myxobacterial vioprolides. Anisomycin, aciclovir and vioprolides promoted intracellular growth of\n                    <jats:italic>Staphylococcus aureus<\/jats:italic>\n                    , a bacterium that is known to be a target of autophagy. In contrast, anisomycin strongly inhibited influenza A virus and SARS\u2010CoV\u20102 replication. Subsequently, we investigated the influence of these autophagy modulators in a cellular disease model of neuronal vacuolation and spinocerebellar degeneration (NVSD), which is associated with cysteine protease ATG4D mutations. We provide evidence that anisomycin and famciclovir, an aciclovir analogue, can normalise the elevated amount of LC3\u2010positive vesicles in mutant fibroblasts, highlighting their potential for the treatment of NVSD. Thus, the screening method enabled the identification of autophagy\u2010modulating compounds with therapeutic potential.\n                  <\/jats:p>","DOI":"10.1111\/febs.70243","type":"journal-article","created":{"date-parts":[[2025,8,28]],"date-time":"2025-08-28T15:54:04Z","timestamp":1756396444000},"page":"2314-2340","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Newly identified properties of known pharmaceuticals and myxobacterial small molecules revealed by screening for autophagy modulators"],"prefix":"10.1111","volume":"293","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-4732-0839","authenticated-orcid":false,"given":"Janine","family":"Fichtner","sequence":"first","affiliation":[{"name":"Systems\u2010Oriented Immunology and Inflammation Research Group Helmholtz Centre for Infection Research  Braunschweig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3236-4362","authenticated-orcid":false,"given":"Yan Yan","family":"Beer","sequence":"additional","affiliation":[{"name":"Systems\u2010Oriented Immunology and Inflammation Research Group Helmholtz Centre for Infection Research  Braunschweig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8456-3246","authenticated-orcid":false,"given":"H. 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