Garcia Gene L, Rericha Erin C, Heger Christopher D, Goldsmith Paul K, Parent Carole A
Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Mol Biol Cell. 2009 Jul;20(14):3295-304. doi: 10.1091/mbc.e09-03-0223. Epub 2009 May 28.
Starvation of Dictyostelium induces a developmental program in which cells form an aggregate that eventually differentiates into a multicellular structure. The aggregate formation is mediated by directional migration of individual cells that quickly transition to group migration in which cells align in a head-to-tail manner to form streams. Cyclic AMP acts as a chemoattractant and its production, secretion, and degradation are highly regulated. A key protein is the extracellular phosphodiesterase PdsA. In this study we examine the role and localization of PdsA during chemotaxis and streaming. We find that pdsA(-) cells respond chemotactically to a narrower range of chemoattractant concentrations compared with wild-type (WT) cells. Moreover, unlike WT cells, pdsA(-) cells do not form streams at low cell densities and form unusual thick and transient streams at high cell densities. We find that the intracellular pool of PdsA is localized to the endoplasmic reticulum, which may provide a compartment for storage and secretion of PdsA. Because we find that cAMP synthesis is normal in cells lacking PdsA, we conclude that signal degradation regulates the external cAMP gradient field generation and that the group migration behavior of these cells is compromised even though their signaling machinery is intact.
盘基网柄菌的饥饿诱导了一个发育程序,在此程序中细胞形成聚集体,最终分化为多细胞结构。聚集体的形成由单个细胞的定向迁移介导,这些细胞会迅速转变为群体迁移,细胞以头对尾的方式排列形成流。环磷酸腺苷(cAMP)作为一种趋化剂,其产生、分泌和降解受到高度调控。一种关键蛋白是细胞外磷酸二酯酶PdsA。在本研究中,我们研究了PdsA在趋化作用和细胞流形成过程中的作用及定位。我们发现,与野生型(WT)细胞相比,pdsA(-)细胞对趋化剂浓度范围的趋化反应更窄。此外,与WT细胞不同,pdsA(-)细胞在低细胞密度下不形成细胞流,而在高细胞密度下形成异常粗大且短暂的细胞流。我们发现PdsA的细胞内池定位于内质网,这可能为PdsA的储存和分泌提供一个区室。因为我们发现缺乏PdsA的细胞中cAMP合成正常,所以我们得出结论,信号降解调节外部cAMP梯度场的产生,并且即使这些细胞的信号传导机制完好无损,它们的群体迁移行为也会受到损害。