A large-scale screen reveals genes that mediate electrotaxis in Dictyostelium discoideum.
A large-scale screen reveals genes that mediate electrotaxis in Dictyostelium discoideum.
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DOI：
10.1126/scisignal.aab0562
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发表时间：
2015-05-26
影响因子：
7.3
通讯作者：
Zhao M
中科院分区：
文献类型：
--
作者：
Gao R;Zhao S;Jiang X;Sun Y;Zhao S;Gao J;Borleis J;Willard S;Tang M;Cai H;Kamimura Y;Huang Y;Jiang J;Huang Z;Mogilner A;Pan T;Devreotes PN;Zhao M
细胞在电场中的定向迁移，一种称为趋流或趋电的现象，发生在许多类型的细胞中，并且可能在伤口愈合和发展中起重要作用。小的细胞外电场可以引导变形虫细胞的迁移，在这里，我们建立了一种大规模的筛选方法，从563株具有形态缺陷的盘状盘形盘形齿柱菌株中寻找具有趋电表型的突变体。我们鉴定出28株有电致性缺陷，10株有稍高的定向响应。通过质粒修复和基因破坏，我们发现了一些突变基因，包括一些先前与趋化性有关的基因。其中，我们研究了PiaA，它编码TORC2的一个关键成分，TORC2是一种激酶蛋白复合物，通过激活激酶PKB（也称为Akt）来传导运动的变化。此外，我们发现缺乏gefA、rasC、rip3、lst8或pkbR1这些编码TORC2-PKB通路其他组分的基因的突变体的趋电性降低。因此，我们开发了一种高通量筛选技术，这将是阐明电趋向性分子机制的有用工具。
Directional cell migration in an electric field, a phenomenon called galvanotaxis or electrotaxis, occurs in many types of cells, and may play an important role in wound healing and development. Small extracellular electric fields can guide the migration of amoeboid cells, and here, we established a large-scale screening approach to search for mutants with electrotaxis phenotypes from a collection of 563 Dictyostelium discoideum strains with morphological defects. We identified 28 strains that were defective in electrotaxis and 10 strains with a slightly higher directional response. Using plasmid rescue followed by gene disruption, we identified some of the mutated genes, including some previously implicated in chemotaxis. Amongst these we studied PiaA, which encodes a critical component of TORC2, a kinase protein complex that transduces changes in motility by activating the kinase PKB (also known as Akt). Furthermore, we found that electrotaxis was decreased in mutants lacking gefA, rasC, rip3, lst8 or pkbR1, genes that encode other components of the TORC2-PKB pathway. Thus, we have developed a high-throughput screening technique that will be a useful tool to elucidate the molecular mechanisms of electrotaxis.
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DOI：
10.1085/jgp.69.6.743
发表时间：
1977-06
期刊：
The Journal of general physiology
影响因子：
--
作者：
Nuccitelli R;Poo MM;Jaffe LF
通讯作者：
Jaffe LF
影响因子：
4
作者：
Graham, David M.;Huang, Ling;Messerli, Mark A.
通讯作者：
Messerli, Mark A.
影响因子：
9.2
作者：
Allen, Greg M.;Mogilner, Alex;Theriot, Julie A.
通讯作者：
Theriot, Julie A.
影响因子：
2.9
作者：
Nuccitelli, Richard;Nuccitelli, Pamela;Smith, Peter J. S.
通讯作者：
Smith, Peter J. S.
DOI：
10.1073/pnas.89.18.8803
发表时间：
1992-09-15
影响因子：
11.1
作者：
KUSPA, A;LOOMIS, WF
通讯作者：
LOOMIS, WF