Studies of the roles of ADP-ribosylation factors and phospholipase D in phorbol ester-induced membrane ruffling
Studies of the roles of ADP-ribosylation factors and phospholipase D in phorbol ester-induced membrane ruffling
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DOI：
10.1002/jcp.20156
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发表时间：
2005-02-01
影响因子：
5.6
通讯作者：
Exton, JH
中科院分区：
文献类型：
--
作者：
Hiroyama, M;Exton, JH
在这项研究中，我们探讨了ADP核糖基化因子（ARF），磷脂酶D（PLD）同工酶，和arfaptins在佛波酯（PMA）诱导的HeLa细胞膜皱褶的作用。PMA刺激诱导皱裂和易位coronin质膜。显性阴性（DN）-ARF 6，DN-ARF 1和DN-Rac 1抑制coronin易位，但不受DN-RhoA和DN-Cdc 42的抑制。不能DN形式的ARF 6，ARF 1，和Rac 1的影响PLD活性响应PMA表明，这种酶没有通过这些小G蛋白激活，它的激活是不是必不可少的诱导皱褶。PMA刺激后，内源性-ARF 1、-ARF 6和-Rac 1与coronin一起沿着存在于皱褶区。DN-ARF 1对DA-ARF 6和DA-Rac 1诱导的皱波无影响，DN-ARF 6对DA-ARF 1和DA-Rac 1诱导的皱波无影响。另一方面，DN-Rac 1抑制DA-ARF 6的作用，但不抑制DA-ARF 1的作用。这些结果表明，PMA通过ARF 6-Rac 1途径引起膜皱褶，同时也通过ARF 1途径平行运作。PLD 1和PLD 2的过表达抑制PMA诱导的corrugin易位和肌动蛋白corrugin复合物的形成，支持这一观点，这些酶是不需要的ruffling，但实际上抑制it. We的结论是PMA诱导的膜ruffling是通过ARF 6-Rac 1和ARF 1途径平行运作，PLD可能抑制。(C)2004 Wiley-Liss，Inc.
In this study, we have explored the roles of ADP-ribosylation factors (ARFs), phospholipase D (PLD) isozymes, and arfaptins in phorbol ester (PMA)-induced membrane ruffling in HeLa cells. PMA stimulation induced ruffling and translocated cortactin to the plasma membrane. The cortactin translocation was inhibited by dominant negative (DN)-ARF6, DN-ARF1, and DN-Rac1, but not by DN-RhoA and DN-Cdc42. The inability of DN-forms of ARF6, ARF1, and Rac1 to affect PLD activity in response to PMA indicated that this enzyme was not activated via these small G proteins and that its activation was not essential for the induction of ruffling. Endogenous-ARF1, -ARF6, and -Rac1 existed in the ruffling region along with cortactin after PMA stimulation. DN-ARF1 had no effect on the ruffling induced by DA-ARF6 or DA-Rac1, and DN-ARF6 had no effect on that induced by DA-ARF1 or DA-Rac1. On the other hand DN-Rac1 suppressed the effect of DA-ARF6 but not that of DA-ARF1. These results suggest that PMA causes membrane ruffling via an ARF6-Rac1 pathway and also an ARF1 pathway operating in parallel. Overexpression of PLD1 and PLD2 inhibited PMA-induced cortactin translocation and actin-cortactin complex formation, supporting the view that these enzymes are not required for ruffling, but actually suppress it. We conclude that PMA-induced membrane ruffling is caused via ARF6-Rac1 and ARF1 pathways operating in parallel and that PLD maybe inhibitory. (C) 2004 Wiley-Liss, Inc.