A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant.

A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant.
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DOI:
10.1016/j.jbiotec.2014.08.008
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发表时间:
2014-12-10
影响因子:
4.1
通讯作者:
Gaertner, Wolfgang
Gaertner, Wolfgang
中科院分区:
工程技术3区
文献类型:
--
作者:
Chen, Zhi-hui;Raffelberg, Sarah;Losi, Aba;Schaap, Pauline;Gaertner, Wolfgang

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先前从蓝藻微鞘藻 PCC7420 中鉴定出光调节腺苷酸环化酶 mPAC。 MPAC 由黄素基蓝光传感 LOV 结构域和催化结构域组成。在这项工作中,我们在真核生物盘基网柄菌的腺苷酸环化酶 A 无效突变体 (aca-) 中表达 mPAC,并测试 mPAC 的光激活可以在多大程度上恢复该生物体的 cAMP 依赖性发育程序。盘基网柄菌属的变形虫是一种成熟的模式生物,它产生并响应 cAMP 脉冲,导致它们聚集并构建子实体。 mPAC 在盘状 D. 中在组成型肌动蛋白 15 启动子的控制下表达,并在黑暗中表现出较低的基础腺苷酸环化酶活性,该活性约为蓝光刺激的五倍。 mPAC 在金合欢细胞中的表达略微恢复了黑暗中的聚集和子实体形成。然而,当表达 mPAC 的细胞在光下孵育时,会形成更多更大的子实体。这些结果扩展了光调节 AC 的先前应用,表明 mPAC 可用于以光依赖性方式操纵真核生物中的多细胞发育。
A light-regulated adenylyl cyclase, mPAC, was previously identified from the cyanobacterium Microcoleus chthonoplastes PCC7420. MPAC consists of a flavin-based blue light-sensing LOV domain and a catalytic domain. In this work, we expressed mPAC in an adenylate cyclase A null mutant (aca-) of the eukaryote Dictyostelium discoideum and tested to what extent light activation of mPAC could restore the cAMP-dependent developmental programme of this organism. Amoebas of Dictyostelium, a well-established model organism, generate and respond to cAMP pulses, which cause them to aggregate and construct fruiting bodies. mPAC was expressed under control of a constitutive actin-15 promoter in D. discoideum and displayed low basal adenylyl cyclase activity in darkness that was about five-fold stimulated by blue light. mPAC expression in aca- cells marginally restored aggregation and fruiting body formation in darkness. However, more and larger fruiting bodies were formed when mPAC expressing cells were incubated in light. Extending former applications of light-regulated AC, these results demonstrate that mPAC can be used to manipulate multicellular development in eukaryotes in a light dependent manner.
腺苷酸环化酶R的功能解剖,这是孢子封装的诱导剂。
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影响因子: --
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