Identification of a common secondary mutation in the <i>Neurospora crassa</i> knockout collection conferring a cell fusion-defective phenotype

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dc.contributor.authorMontenegro-Montero, Alejandro
dc.contributor.authorGoity, Alejandra
dc.contributor.authorCanessa, Paulo F.
dc.contributor.authorLarrondo, Luis F.
dc.date.accessioned2025-01-20T17:21:57Z
dc.date.available2025-01-20T17:21:57Z
dc.date.issued2023
dc.description.abstractGene-deletion mutants represent a powerful tool to study gene function. The filamentous fungus Neurospora crassa is a well-established model organism, and features a comprehensive gene knockout strain collection. While these mutant strains have been used in numerous studies, resulting in the functional annotation of many Neurospora genes, direct confirmation of gene-phenotype relationships is often lacking, which is particularly relevant given the possibility of background mutations, sample contamination, and/or strain mislabeling. Indeed, spontaneous mutations resulting in phenotypes resembling many cell fusion mutants have long been known to occur at relatively high frequency in N. crassa, and these secondary mutations are common in the Neurospora deletion collection. The identity of these mutations, however, is largely unknown. Here, we report that the Delta ada-3 strain from the N. crassa knockout collection, which exhibits a cell fusion defect, harbors a secondary mutation responsible for this phenotype. Through whole-genome sequencing and genetic analyses, we found a similar to 30-Kb deletion in this strain affecting a known cell fusion-related gene, so/ham-1, and show that it is the absence of this gene-and not of ada-3-that underlies its cell fusion defect. We additionally found three other knockout strains harboring the same deletion, suggesting that this mutation may be common in the collection and could have impacted previous studies. Our findings provide a cautionary note and highlight the importance of proper functional validation of strains from mutant collections. We discuss our results in the context of the spread of cell fusion-defective cheater variants in N. crassa cultures.
dc.description.abstractIMPORTANCE This study emphasizes the need for careful and detailed characterization of strains from mutant collections. Specifically, we found a common deletion in various strains from the Neurospora crassa gene knockout collection that results in a cell fusion-defective phenotype. This is noteworthy because this collection is known to contain background mutations-of a largely unclear nature-that produce cell fusion-defective phenotypes. Our results describe an example of such mutations, and highlight how this common genetic defect could have impacted previous studies that have used the affected strains. Furthermore, they provide a cautionary note about the use of Neurospora strains with similar phenotypes. Lastly, these findings offer additional details relevant to our understanding of the origin and spread of cell fusion-defective cheater variants in N. crassa cultures.
dc.fuente.origenWOS
dc.identifier.doi10.1128/spectrum.02087-23
dc.identifier.issn2165-0497
dc.identifier.urihttps://doi.org/10.1128/spectrum.02087-23
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/91471
dc.identifier.wosidWOS:001107303900167
dc.issue.numero5
dc.language.isoen
dc.revistaMicrobiology spectrum
dc.rightsacceso restringido
dc.subjectNeurospora crassa
dc.subjectcell fusion
dc.subjectgene knockout
dc.subjectsecondary mutations
dc.titleIdentification of a common secondary mutation in the <i>Neurospora crassa</i> knockout collection conferring a cell fusion-defective phenotype
dc.typeartículo
dc.volumen11
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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