{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:53:49Z","timestamp":1791294829672,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32400477"],"award-info":[{"award-number":["32400477"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Metastable phenotypic inheritance is often attributed to epigenetic mechanisms, but reversible genetic alterations can produce similar instability. Here, we investigated the basis of unstable resistance to TORC1 inhibitor (rapamycin plus caffeine) in\n                      <jats:italic toggle=\"yes\">Schizosaccharomyces pombe<\/jats:italic>\n                      . Six independent, metastable resistant mutants were isolated. Genetic mapping positioned the causal lesion to a single Mendelian locus, which sequencing identified as\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      , encoding a GATA transcription factor and a key negative regulator of growth downstream of TORC1. In each mutant, distinct loss-of-function mutations (insertions, deletions, or point mutations) were found in\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      in the resistant state, and these mutations precisely reverted to the wild-type sequence upon loss of resistance. Restoring the wild-type\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      allele abolished resistance, indicating that reversible genetic disruption of\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      is both necessary and sufficient for the metastable phenotype. Furthermore, strong resistance in several strains from a genome-wide deletion library was due to secondary, inactivating mutations in\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      , underscoring its role as a recurrent adaptive target under rapamycin plus caffeine treatment. Mechanistically,\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      inactivation established a distinct basal transcriptome and pronounced derepression of translation and metabolic programs upon drug treatment. While rapamycin plus caffeine triggered extensive chromatin remodeling and H3K9 methylation contributed partially to resistance, these epigenetic changes were most consistent with a downstream modifying layer. Our study shows that metastable drug resistance in fission yeast is predominantly associated with recurrent, reversible genetic inactivation of the central transcriptional regulator\n                      <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                      , demonstrating how rapidly reversible genetic switches can drive adaptive evolution.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Distinguishing between genetic and epigenetic inheritance is fundamental to understanding how cells adapt to environmental stress. In the fission yeast\n                        <jats:italic toggle=\"yes\">Schizosaccharomyces pombe<\/jats:italic>\n                        , rapid and reversible drug resistance is often assumed to be driven by epigenetic switches that change gene activity without altering DNA. However, our study reveals that this instability can be caused by physical mutations in a single gene,\n                        <jats:italic toggle=\"yes\">gaf1<\/jats:italic>\n                        , which acts as a genetic toggle. These mutations appear under drug pressure and precisely revert to the original sequence when the drug is removed. We also demonstrate that these spontaneous mutations can contaminate standard laboratory yeast collections, leading to potential misinterpretation of experimental data. These findings broaden our understanding of unstable inheritance and show that DNA sequences can be far more dynamic than previously recognized during rapid evolution and the development of drug resistance.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01467-26","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T13:03:56Z","timestamp":1789650236000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Recurrent reversible mutations at\n                    <i>gaf1<\/i>\n                    driving metastable TORC1 inhibitor resistance in fission yeast"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7477-7459","authenticated-orcid":true,"given":"Lufeng","family":"Dan","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/034z67559","id-type":"ROR","asserted-by":"publisher"}],"name":"Antibiotics Innovation and Resistance Control Key Laboratory of Sichuan Province, School of Pharmacy, Chengdu University","place":["Chengdu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaowen","family":"Ye","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/034z67559","id-type":"ROR","asserted-by":"publisher"}],"name":"Antibiotics Innovation and Resistance Control Key Laboratory of Sichuan Province, School of Pharmacy, Chengdu University","place":["Chengdu, 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