Overexpression of Poly(ADP-ribose) Polymerase Disrupts Organization of Cytoskeletal F-actin and Tissue Polarity inDrosophila *
Overexpression of Poly(ADP-ribose) Polymerase Disrupts Organization of Cytoskeletal F-actin and Tissue Polarity inDrosophila *
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DOI：
10.1074/jbc.m109826200
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发表时间：
2002-02
期刊：
影响因子：
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通讯作者：
M. Uchida;S. Hanai;N. Uematsu;K. Sawamoto;H. Okano;M. Miwa;K. Uchida
中科院分区：
文献类型：
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作者：
M. Uchida;S. Hanai;N. Uematsu;K. Sawamoto;H. Okano;M. Miwa;K. Uchida
聚（ADP-核糖）聚合酶（PARP）可能在细胞周期、细胞增殖和染色体稳定性维持等核事件中发挥重要作用。然而，PARP 所发挥的确切生物学作用或 PARP 如何参与这些细胞功能仍不清楚。为了阐明 PARP 在体内的生物学功能，我们构建了在发育中的眼原基中过度表达果蝇 PARP 的转基因果蝇。这些果蝇表现出正常光滑的小眼晶格的轻度粗糙化和由于小眼旋转和手性不当引起的组织极性破坏。为了阐明这种表型变化是如何诱导的，我们在这里详细分析了在发育中的眼睛、胚胎和成虫中过度表达 PARP 的转基因果蝇。携带更多转基因拷贝的果蝇中 PARP mRNA 水平和表型得到增强。通过使用神经细胞标记来分析第三龄幼虫发育中的眼睛，以检查 PARP 在细胞命运中的参与情况。在 PARP 转基因果蝇中观察到非神经元辅助细胞的形态紊乱。有趣的是，PARP的过度表达不会干扰细胞周期或细胞凋亡，但它确实破坏了细胞骨架F-肌动蛋白的组织，导致细胞和组织形态异常。此外，热诱导的 PARP 表达破坏了胚胎中细胞骨架 F-肌动蛋白的组织和成年果蝇的组织极性。由于这些表型与组织极性基因的突变体或转基因果蝇非常相似，因此检查了 PARP 与已知组织极性基因的遗传相互作用。将眼睛中表达 PARP 或 RhoA GTPase 的转基因果蝇进行杂交，PARP 的共表达抑制了 RhoA GTPase 的作用。我们的结果表明 PARP 可能在果蝇发育过程中的细胞骨架或细胞质事件中发挥作用。
Poly(ADP-ribose) polymerase (PARP) may play important roles in nuclear events such as cell cycle, cell proliferation, and maintenance of chromosomal stability. However, the exact biological role played by PARP or how PARP is involved in these cellular functions is still unclear. To elucidate the biological functions of PARP in vivo, we have constructed transgenic flies that overexpress Drosophila PARP in the developing eye primordia. These flies showed mild roughening of the normally smooth ommatidial lattice and tissue polarity disruption caused by improper rotation and chirality of the ommatidia. To clarify how this phenotypical change was induced, here we analyzed transgenic flies overexpressing PARP in the developing eye, embryo, and adult in detail. PARP mRNA level and the phenotype were enhanced in flies carrying more copies of the transgene. Developing eyes from third instar larvae were analyzed by using the neural cell marker to examine the involvement of PARP in cell fate. Morphological disorder of non-neuronal accessory cells was observed in PARP transgenic flies. Interestingly, overexpression of PARP did not interfere with the cell cycle or apoptosis, but it did disrupt the organization of cytoskeletal F-actin, resulting in aberrant cell and tissue morphology. Furthermore, heat-induced PARP expression disrupted organization of cytoskeletal F-actin in embryos and tissue polarity in adult flies. Because these phenotypes closely resembled mutants or transgenic flies of the tissue polarity genes, genetic interaction of PARP with known tissue polarity genes was examined. Transgenic flies expressing either PARP or RhoA GTPase in the eye were crossed, and co-expression of PARP suppressed the effect of RhoA GTPase. Our results indicate that PARP may play a role in cytoskeletal or cytoplasmic events in developmental processes of Drosophila.