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Polarized gene expression determines Woronin body formation at the leading edge of the fungal colony

机译:极化的基因表达决定了真菌菌落前沿的沃罗宁体形成

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The Woronin body (WB) is a peroxisome-related organelle that is centered on a crystalline core of the HEX-1 protein, which functions to seal septal pores of filamentous ascomycetes in response to cellular damage. Here, we investigate the cellular and genetic control of WB-formation and show that polarized hex-1 gene expression determines WB-biogenesis at the growing hyphal apex. We find that intron splicing is coupled to efficient hex-1 gene expression and strikingly, when the yellow fluorescent protein was expressed from hex-1 regulatory sequences, we observed a fluorescent gradient that was maximal in apical cells. Moreover, endogenous hex-1 transcripts were specifically enriched at the leading edge of the fungal colony, whereas other transcripts accumulated in basal regions. Time-lapse confocal microscopy showed that HEX-1 crystals normally formed in the vicinity of the hyphal apex in large peroxisomes, which matured and were immobilized at the cell periphery as cells underwent septation. When the hex-1 structural gene was expressed from regulatory sequences of an abundant, basally localized transcript, WB-core formation was redetermined to basal regions of the colony, and these strains displayed loss-of-function phenotypes specifically in apical hyphal compartments. These results show that apically localized gene expression is a key determinant of spatially restricted WB-assembly. We suggest that this type of regulation may be widely used to determine cellular activity in apical regions of the fungal hypha.
机译:Woronin体(WB)是与过氧化物酶体有关的细胞器,其中心位于HEX-1蛋白的结晶核心上,可响应细胞损伤而封闭丝状子囊的间隔孔。在这里,我们调查的WB形成的细胞和遗传控制,并表明极化的hex-1基因表达决定了在生长的菌丝顶点处的WB生物发生。我们发现内含子剪接与有效的hex-1基因表达耦合,并且惊人地,当从hex-1调控序列表达黄色荧光蛋白时,我们观察到在顶端细胞中最大的荧光梯度。此外,内源性hex-1转录物在真菌菌落的前缘特别富集,而其他转录物则在基底区域积累。延时共聚焦显微镜显示,HEX-1晶体通常在大型过氧化物酶体的菌丝顶点附近形成,随着细胞的分离成熟并固定在细胞外围。当hex-1结构基因从丰富的,基本局部的转录物的调控序列中表达时,WB核心的形成被重新确定到菌落的基础区域,并且这些菌株显示出功能丧失的表型,特别是在根尖菌丝区室。这些结果表明,顶端定位的基因表达是空间受限的WB组装的关键决定因素。我们建议这种类型的监管可能被广泛用于确定真菌菌丝的顶端区域中的细胞活性。

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