重新评估果蝇中 Bazooka/Par-3 的亚细胞定位:没有证据表明其定位在细胞核和神经肌肉接头处。

IF 1.8 4区 生物学 Q3 BIOLOGY Biology Open Pub Date : 2024-06-15 Epub Date: 2024-06-27 DOI:10.1242/bio.060544
Soya Kim, Jaffer Shahab, Elisabeth Vogelsang, Andreas Wodarz
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引用次数: 0

摘要

Bazooka/Par-3(Baz)是一种进化保守的支架蛋白,在许多不同类型的细胞中起着建立和维持细胞极性的主调控因子的作用。据报道,在绝大多数已发表的研究论文中,Baz 定位于细胞皮层和细胞间连接处。不过,也有一些报告显示 Baz 在其他亚细胞位点的定位和功能,特别是在核膜和神经肌肉接头处。在本研究中,我们以系统的方式重新评估了 Baz 在这些亚细胞位点的定位。我们使用在不同宿主动物体内针对 Baz 不同表位制备的抗体对果蝇组织进行共聚焦成像。我们通过对baz等位基因空突变体的镶嵌分析和针对baz的组织特异性RNAi测试了这些抗血清的特异性。此外,我们还使用了Baz的GFP标记基因诱捕系和细菌人工染色体(BAC),在baz突变体背景下,在其内源启动子的控制下表达GFP标记的Baz,以比较GFP-Baz融合蛋白的亚细胞定位与抗Baz抗血清的染色。总之,这些实验没有提供证据证明 Baz 在细胞核或神经肌肉接头处的特异性定位。
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Re-assessment of the subcellular localization of Bazooka/Par-3 in Drosophila: no evidence for localization to the nucleus and the neuromuscular junction.

Bazooka/Par-3 (Baz) is an evolutionarily conserved scaffold protein that functions as a master regulator for the establishment and maintenance of cell polarity in many different cell types. In the vast majority of published research papers Baz has been reported to localize at the cell cortex and at intercellular junctions. However, there have also been several reports showing localization and function of Baz at additional subcellular sites, in particular the nuclear envelope and the neuromuscular junction. In this study we have re-assessed the localization of Baz to these subcellular sites in a systematic manner. We used antibodies raised in different host animals against different epitopes of Baz for confocal imaging of Drosophila tissues. We tested the specificity of these antisera by mosaic analysis with null mutant baz alleles and tissue-specific RNAi against baz. In addition, we used a GFP-tagged gene trap line for Baz and a bacterial artificial chromosome (BAC) expressing GFP-tagged Baz under control of its endogenous promoter in a baz mutant background to compare the subcellular localization of the GFP-Baz fusion proteins to the staining with anti-Baz antisera. Together, these experiments did not provide evidence for specific localization of Baz to the nucleus or the neuromuscular junction.

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来源期刊
Biology Open
Biology Open BIOLOGY-
CiteScore
3.90
自引率
0.00%
发文量
162
审稿时长
8 weeks
期刊介绍: Biology Open (BiO) is an online Open Access journal that publishes peer-reviewed original research across all aspects of the biological sciences. BiO aims to provide rapid publication for scientifically sound observations and valid conclusions, without a requirement for perceived impact.
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