EB-SUN, a New Microtubule Plus-End Tracking Protein in Drosophila.

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

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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EB-SUN--果蝇中一种新的微管加端追踪蛋白
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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