Rodriguez-Centeno Javier, Sastre Leandro
Instituto de Investigaciones Biomédicas CSIC/UAM, C/Arturo Duperier, 4, 28029, Madrid, Spain.
CIBERER, Valencia, Spain.
PLoS One. 2016 Feb 3;11(2):e0148533. doi: 10.1371/journal.pone.0148533. eCollection 2016.
Amoebae of the Dictyostelium discoideum species form multicellular fruiting bodies upon starvation. Cyclic adenosine monophosphate (cAMP) is used as intercellular signalling molecule in cell-aggregation, cell differentiation and morphogenesis. This molecule is synthesized by three adenylyl cyclases, one of which, ACA, is required for cell aggregation. The gene coding for ACA (acaA) is transcribed from three different promoters that are active at different developmental stages. Promoter 1 is active during cell-aggregation, promoters 2 and 3 are active in prespore and prestalk tip cells at subsequent developmental stages. The biological relevance of acaA expression from each of the promoters has been studied in this article. The acaA gene was expressed in acaA-mutant cells, that do not aggregate, under control of each of the three acaA promoters. acaA expression under promoter 1 control induced cell aggregation although subsequent development was delayed, very small fruiting bodies were formed and cell differentiation genes were expressed at very low levels. Promoter 2-driven acaA expression induced the formation of small aggregates and small fruiting bodies were formed at the same time as in wild-type strains and differentiation genes were also expressed at lower levels. Expression of acaA from promoter 3 induced aggregates and fruiting bodies formation and their size and the expression of differentiation genes were more similar to that of wild-type cells. Expression of acaA from promoters 1 and 2 in AX4 cells also produced smaller structures. In conclusion, the expression of acaA under control of the aggregation-specific Promoter 1 is able to induce cell aggregation in acaA-mutant strains. Expression from promoters 2 and 3 also recovered aggregation and development although promoter 3 induced a more complete recovery of fruiting body formation.
盘基网柄菌属的变形虫在饥饿时会形成多细胞子实体。环磷酸腺苷(cAMP)在细胞聚集、细胞分化和形态发生过程中用作细胞间信号分子。该分子由三种腺苷酸环化酶合成,其中一种,即ACA,是细胞聚集所必需的。编码ACA的基因(acaA)从三个不同的启动子转录,这些启动子在不同的发育阶段具有活性。启动子1在细胞聚集期间具有活性,启动子2和3在随后发育阶段的前孢子和前柄顶端细胞中具有活性。本文研究了每个启动子驱动的acaA表达的生物学意义。在三个acaA启动子各自的控制下,acaA基因在不发生聚集的acaA突变细胞中表达。在启动子1控制下的acaA表达诱导了细胞聚集,尽管随后的发育延迟,形成的子实体非常小,并且细胞分化基因的表达水平非常低。启动子2驱动的acaA表达诱导形成小聚集体,同时形成的小果实与野生型菌株中的情况相同,分化基因也以较低水平表达。启动子3驱动的acaA表达诱导了聚集体和子实体的形成,它们的大小和分化基因的表达与野生型细胞更相似。在AX4细胞中,启动子1和2驱动的acaA表达也产生了较小的结构。总之,在聚集特异性启动子1控制下的acaA表达能够在acaA突变菌株中诱导细胞聚集。启动子2和3的表达也恢复了聚集和发育,尽管启动子3诱导子实体形成的恢复更为完全。