Dorighetto Cogo Antônio Jesus, Dutra Ferreira Keilla Dos Reis, Okorokov Lev A, Ramos Alessandro C, Façanha Arnoldo R, Okorokova-Façanha Anna L
Laboratório de Fisiologia e Bioquímica de Microrganismos, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego, 2000, Pq. Califórnia, Campos dos Goytacazes-RJ 28013-602, Brazil.
Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego, 2000, Pq. Califórnia, Campos dos Goytacazes-RJ 28013-602, Brazil.
Biol Open. 2018 Feb 20;7(2):bio029660. doi: 10.1242/bio.029660.
Polyamines play a regulatory role in eukaryotic cell growth and morphogenesis. Despite many molecular advances, the underlying mechanism of action remains unclear. Here, we investigate a mechanism by which spermine affects the morphogenesis of a dimorphic fungal model of emerging relevance in plant interactions, , through the recruitment of a phytohormone-like pathway involving activation of the plasma membrane P-type H-ATPase. Morphological transition was followed microscopically, and the H-ATPase activity was analyzed in isolated membrane vesicles. Proton flux and acidification were directly probed at living cell surfaces by a non-invasive selective ion electrode technique. Spermine and indol-3-acetic acid (IAA) induced the yeast-hypha transition, influencing the colony architecture. Spermine induced H-ATPase activity and H efflux in living cells correlating with yeast-hypha dynamics. Pharmacological inhibition of spermine and IAA pathways prevented the physio-morphological responses, and indicated that spermine could act upstream of the IAA pathway. This study provides the first compelling evidence on the fungal morphogenesis and colony development as modulated by a spermine-induced acid growth mechanism analogous to that previously postulated for the multicellular growth regulation of plants.
多胺在真核细胞生长和形态发生中发挥调节作用。尽管在分子层面取得了许多进展,但其潜在作用机制仍不清楚。在此,我们研究了一种机制,即精胺通过招募一种涉及质膜P型H⁺ -ATP酶激活的类植物激素途径,来影响一种在植物相互作用中新兴的双态真菌模型的形态发生。通过显微镜观察形态转变,并在分离的膜囊泡中分析H⁺ -ATP酶活性。利用非侵入性选择性离子电极技术直接在活细胞表面探测质子通量和酸化情况。精胺和吲哚 - 3 - 乙酸(IAA)诱导酵母 - 菌丝转变,影响菌落结构。精胺诱导活细胞中的H⁺ -ATP酶活性和H⁺ 外流,这与酵母 - 菌丝动态变化相关。对精胺和IAA途径的药理学抑制阻止了生理形态反应,并表明精胺可能在IAA途径的上游起作用。本研究首次提供了令人信服的证据,证明精胺诱导的酸生长机制对真菌形态发生和菌落发育的调节作用,该机制类似于先前假定的植物多细胞生长调节机制。