Stowers Institute for Medical Research, Kansas City, Missouri, United States of America.
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
PLoS Genet. 2022 Oct 27;18(10):e1010462. doi: 10.1371/journal.pgen.1010462. eCollection 2022 Oct.
Numerous genes required for sexual reproduction remain to be identified even in simple model species like Schizosaccharomyces pombe. To address this, we developed an assay in S. pombe that couples transposon mutagenesis with high-throughput sequencing (TN-seq) to quantitatively measure the fitness contribution of nonessential genes across the genome to sexual reproduction. This approach identified 532 genes that contribute to sex, including more than 200 that were not previously annotated to be involved in the process, of which more than 150 have orthologs in vertebrates. Among our verified hits was an uncharacterized gene, ifs1 (important for sex), that is required for spore viability. In two other hits, plb1 and alg9, we observed a novel mutant phenotype of poor spore health wherein viable spores are produced, but the spores exhibit low fitness and are rapidly outcompeted by wild type. Finally, we fortuitously discovered that a gene previously thought to be essential, sdg1 (social distancing gene), is instead required for growth at low cell densities and can be rescued by conditioned medium. Our assay will be valuable in further studies of sexual reproduction in S. pombe and identifies multiple candidate genes that could contribute to sexual reproduction in other eukaryotes, including humans.
即使在像裂殖酵母这样的简单模式物种中，仍有许多参与有性生殖的基因有待鉴定。为了解决这个问题，我们在裂殖酵母中开发了一种测定方法，该方法将转座子诱变与高通量测序（TN-seq）相结合，定量测量基因组中非必需基因对有性生殖的适应性贡献。这种方法鉴定出了 532 个对性有贡献的基因，其中包括 200 多个以前未注释为参与该过程的基因，其中超过 150 个在脊椎动物中有同源基因。在我们验证的命中中，有一个未被描述的基因 ifs1（对性别很重要），它是孢子活力所必需的。在另外两个命中 plb1 和 alg9 中，我们观察到一种新的孢子健康不良的突变表型，其中可产生有活力的孢子，但孢子的适应性差，很快就被野生型所淘汰。最后，我们偶然发现了一个以前被认为是必需的基因 sdg1（社交距离基因），它实际上是在低细胞密度下生长所必需的，可以通过条件培养基来挽救。我们的测定方法将在裂殖酵母有性生殖的进一步研究中具有重要价值，并鉴定出多个候选基因，这些基因可能对其他真核生物（包括人类）的有性生殖有贡献。
