A novel partner for Dictyostelium filamin is an α-helical developmentally regulated protein
A novel partner for Dictyostelium filamin is an α-helical developmentally regulated protein
复制标题
盘基网柄菌丝蛋白的新伙伴是一种 α 螺旋发育调节蛋白
作者：
M. Knuth;N. Khaire;A. Kuspa;Sijie Lu;M. Schleicher;A. Noegel
丝蛋白是一个高度同源的肌动蛋白交联蛋白家族，它稳定了三维肌动蛋白网络，将它们连接到膜蛋白上，并通过与各种结合伙伴的相互作用将细胞内的信号反应引导到肌动蛋白支架上。在这里，我们描述了第一个分离到的盘状钢丝蛋白相互作用蛋白- FIP，一个229.8 kDa的蛋白，具有两个α-螺旋卷曲的线圈结构域。以丝蛋白棒状结构域为诱饵，在酵母双杂交筛选中鉴定出FIP。FIP也可以与细胞提取物中的丝蛋白共免疫沉淀。缺失分析将FIP相互作用域定位在c端区域；相反，在丝蛋白棒中，重复序列2-4与重组FIP蛋白相互作用。7 kb的FIP转录本在发育早期被上调。针对细菌表达的FIP多肽提出的单克隆抗体在western blots中识别230 kDa的发育调节蛋白。免疫荧光分析显示细胞质中有点状染色，在细胞分离实验中，FIP主要存在于细胞质部分。一种由GFP和c端组成的融合蛋白定位于质膜并与细胞骨架相关。融合蛋白的表达影响发育，影响多细胞聚集体的大小和鼻涕虫的趋光行为。因此，FIP可能在动态肌动蛋白细胞骨架和阿米巴Dictyostelium amoebae多细胞阶段的信号转导事件之间提供了一个候选的联系。
The filamins are a family of highly homologous actincrosslinking proteins that stabilize three-dimensional actin networks, link them to membrane proteins and direct intracellular signaling reactions to the actin scaffold through interaction with various binding partners. Here, we describe the first Dictyostelium filamin-interacting protein to be isolated - FIP, a 229.8 kDa protein with two α-helical coiled coil domains. FIP was identified in a yeast two-hybrid screen using the rod domain of filamin as bait. FIP can also be coimmunoprecipitated with filamin from cellular extracts. Deletion analysis located the interaction domain of FIP to a C-terminal region; by contrast, in filamin rods, repeats 2-4 interacted with the recombinant FIP protein. The 7 kb transcript of FIP is upregulated during early development. Monoclonal antibodies raised against a bacterially expressed FIP polypeptide recognize a 230 kDa developmentally regulated protein in western blots. Immunofluorescence analysis shows a punctate staining pattern in the cytosol and, in cell fractionation experiments, FIP is mainly found in the cytosolic fraction. A fusion protein composed of GFP and the C-terminal part localizes to the plasma membrane and is associated with the cytoskeleton. Expression of the fusion protein affects development and influences the size of the multicellular aggregates and the phototactic behavior of slugs. Thus, FIP might provide a candidate link between the dynamic actin cytoskeleton and signal transduction events during the multicellular stages of Dictyostelium amoebae.
DOI：
10.1073/pnas.82.4.1074
发表时间：
1985-01-01
影响因子：
11.1
作者：
TABOR, S;RICHARDSON, CC
通讯作者：
RICHARDSON, CC
影响因子：
10.5
作者：
DURFEE, T;BECHERER, K;ELLEDGE, SJ
通讯作者：
ELLEDGE, SJ
信号转导突变体中盘基网柄菌早期发育基因的调控。
DOI：
10.1006/dbio.1995.1267
发表时间：
1995
期刊：
Developmental biology.
影响因子：
--
作者：
Wu,L;Hansen,D;Franke,J;Kessin,RH;Podgorski,GJ
通讯作者：
Podgorski,GJ