Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York, United States of America.
PLoS Genet. 2023 Oct 23;19(10):e1011009. doi: 10.1371/journal.pgen.1011009. eCollection 2023 Oct.
The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal, bam genetically interacts with the endosymbiont Wolbachia, as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the wMel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam, suggesting wMel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if wMel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that wMel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of wMel interaction with germline stem cells in bam mutants.
果蝇的 bag of marbles(bam)蛋白编码基因在早期雄性和雌性生殖中起着关键作用,它与伴侣蛋白形成复合物,促进配子发生中的分化。像另一个生殖系基因 Sex lethal 一样,bam 与共生菌 Wolbachia 在遗传上相互作用,因为 Wolbachia 挽救了 bam 弱突变体的生育能力降低。在这里,我们通过生成 22 个新的 bam 突变体来探索 bam-Wolbachia 相互作用的特异性,其中 10 个突变体显示出生育缺陷。其中 9 个突变体倾向于被 wMel Wolbachia 变体拯救,其中 8 个在生育力和/或细胞学水平上具有统计学意义。在某些情况下,生育力显著增加了 20 倍。拯救与 bam 的已知结合区域之间没有特异性,这表明 wMel 不会与一个独特的 bam 伴侣相互作用来拯救生殖表型。我们进一步测试了 wMel 是否以非特异性的方式与 bam 相互作用,方法是增加 bam 转录本水平或在上游作用于生殖干细胞。对 bam RNAi 敲低突变体的生育力评估表明,wMel 拯救是特异性的,针对功能性突变的 bam 等位基因,我们没有发现 wMel 与 bam 突变体中的生殖干细胞相互作用的明显证据。