Rgnef (p190RhoGEF) knockout inhibits RhoA activity, focal adhesion establishment, and cell motility downstream of integrins.
Rgnef (p190RhoGEF) knockout inhibits RhoA activity, focal adhesion establishment, and cell motility downstream of integrins.
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DOI：
10.1371/journal.pone.0037830
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Schlaepfer DD
中科院分区：
文献类型：
--
作者：
Miller NL;Lawson C;Chen XL;Lim ST;Schlaepfer DD
细胞迁移是一个高度调节的过程，涉及称为粘着斑的细胞-基质接触位点的形成和周转。Rho家族GTP酶是调节细胞中肌动蛋白和粘着斑动力学的分子开关。鸟嘌呤核苷酸交换因子（GEF）激活Rho家族GTP酶。Rgnef（p190 RhoGEF）是一种普遍存在的190 kDa GEF，参与结肠癌和成纤维细胞运动的控制。 建立Rgnef外显子24 floxed小鼠（Rgnefflox），并与巨细胞病毒（CMV）驱动的Cre重组酶转基因小鼠杂交，以在早期发育期间抑制Rgnef在所有组织中的表达。杂合子RgnefWT/flox（Cre+）杂交在胚胎第13.5天产生正常的孟德尔比率，但Rgneflox/flox（Cre+）小鼠在3周龄的数量显着低于预期。分离Rgnefflox/flox（Cre+）（Rgnef-/-）胚胎和原代小鼠胚胎成纤维细胞（MEF），并验证其缺乏Rgnef蛋白表达。当与野生型（WT）同窝MEFs相比，Rgnef的损失显着抑制haptotaxis迁移，伤口闭合运动性，粘着斑数，和RhoA GT3激活后，纤连蛋白-整合素刺激。在WT MEF中，在与RhoA活化相关的细胞的纤连蛋白铺板后60分钟内发生Rgnef活化。Rgnef−/− MEF表型通过表位标记的Rgnef再表达得以拯救。Rgnef−/− MEF表型是由于Rgnef丢失，并支持Rgnef在RhoA调节整合素下游控制细胞迁移中的重要作用。
Cell migration is a highly regulated process that involves the formation and turnover of cell-matrix contact sites termed focal adhesions. Rho-family GTPases are molecular switches that regulate actin and focal adhesion dynamics in cells. Guanine nucleotide exchange factors (GEFs) activate Rho-family GTPases. Rgnef (p190RhoGEF) is a ubiquitous 190 kDa GEF implicated in the control of colon carcinoma and fibroblast cell motility. Rgnef exon 24 floxed mice (Rgnefflox) were created and crossed with cytomegalovirus (CMV)-driven Cre recombinase transgenic mice to inactivate Rgnef expression in all tissues during early development. Heterozygous RgnefWT/flox (Cre+) crosses yielded normal Mendelian ratios at embryonic day 13.5, but Rgnefflox/flox (Cre+) mice numbers at 3 weeks of age were significantly less than expected. Rgnefflox/flox (Cre+) (Rgnef−/−) embryos and primary mouse embryo fibroblasts (MEFs) were isolated and verified to lack Rgnef protein expression. When compared to wildtype (WT) littermate MEFs, loss of Rgnef significantly inhibited haptotaxis migration, wound closure motility, focal adhesion number, and RhoA GTPase activation after fibronectin-integrin stimulation. In WT MEFs, Rgnef activation occurs within 60 minutes upon fibronectin plating of cells associated with RhoA activation. Rgnef−/− MEF phenotypes were rescued by epitope-tagged Rgnef re-expression. Rgnef−/− MEF phenotypes were due to Rgnef loss and support an essential role for Rgnef in RhoA regulation downstream of integrins in control of cell migration.
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影响因子：
15.3
作者：
Fujikawa, K;Miletic, AV;Alt, FW;Faccio, R;Brown, T;Hoog, J;Fredericks, J;Nishi, S;Mildiner, S;Moores, SL;Brugge, J;Rosen, FS;Swat, W
通讯作者：
Swat, W
影响因子：
7.2
作者：
Geiger B;Yamada KM
通讯作者：
Yamada KM
影响因子：
6.4
作者：
ENGVALL, E;RUOSLAHTI, E
通讯作者：
RUOSLAHTI, E
影响因子：
9.2
作者：
Bravo-Cordero, Jose Javier;Oser, Matthew;Chen, Xiaoming;Eddy, Robert;Hodgson, Louis;Condeelis, John
通讯作者：
Condeelis, John
影响因子：
7.8
作者：
Gebbink, MFBG;Kranenburg, O;Moolenaar, WH
通讯作者：
Moolenaar, WH