EB-SUN--果蝇中一种新的微管加端追踪蛋白

Sun K Kim, Stephen Rogers, Wen Lu, Brad S. Lee, Vladimir I Gelfand
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摘要

微管(MT)调节对卵母细胞的发育至关重要。在果蝇中,MT的稳定性、极性、丰度和定向在各个发育阶段都会发生动态变化。为了鉴定果蝇卵巢中调控MT的新型微管相关蛋白(MAPs),我们发现了一个之前未表征的基因CG18190,它编码一种新型MT末端结合(EB)蛋白,我们将其命名为EB-SUN。我们发现,在果蝇 S2 细胞中,EB-SUN 与 EB1 共同定位在生长的微管加端。Paralog Explorer 的组织特异性和发育表达谱显示,EB-SUN 主要在卵巢和早期胚胎中表达,而 EB1 则普遍表达。此外,早在卵母细胞形成过程中,EB-SUN 彗星就高度集中在卵母细胞中。EB-SUN 基因敲除(KO)会导致卵子发生中期(第 7 阶段)MT 密度下降,并延迟卵子发生中期晚期(第 9 阶段)的卵母细胞生长。将 EB-SUN KO 与生殖细胞中的 EB1 敲除(KD)相结合,可显著进一步降低第 7 阶段的 MT 密度。值得注意的是,与单基因耗竭不同,EB-SUN KO/EB1 KD 雌性的所有卵子都无法孵化,这表明这两种 EB 蛋白在胚胎发生过程中存在功能冗余。我们的研究结果表明,EB-SUN 和 EB1 在早期胚胎发生过程中发挥着不同的作用。
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EB-SUN, a New Microtubule Plus-End Tracking Protein in Drosophila.
Microtubule (MT) regulation is essential for oocyte development. In Drosophila, MT stability, polarity, abundance, and orientation undergo dynamic changes across developmental stages. In our effort to identify novel microtubule-associated proteins (MAPs) that regulate MTs in the Drosophila ovary, we identified a previously uncharacterized gene, CG18190, encoding a novel MT end-binding (EB) protein, which we propose to name EB-SUN. We show that EB-SUN colocalizes with EB1 at growing microtubule plus-ends in Drosophila S2 cells. Tissue-specific and developmental expression profiles from Paralog Explorer reveal that EB-SUN is predominantly expressed in the ovary and early embryos, while EB1 is ubiquitously expressed. Furthermore, as early as oocyte determination, EB-SUN comets are highly concentrated in oocytes during oogenesis. EB-SUN knockout (KO) results in a decrease in MT density at the onset of mid-oogenesis (Stage 7) and delays oocyte growth during late mid-oogenesis (Stage 9). Combining EB-SUN KO with EB1 knockdown (KD) in germ cells significantly further reduced MT density at Stage 7. Notably, all eggs from EB-SUN KO/EB1 KD females fail to hatch, unlike single gene depletion, suggesting a functional redundancy between these two EB proteins during embryogenesis. Our findings indicate that EB-SUN and EB1 play distinct roles during early embryogenesis.
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