Garrido-Bazán Verónica, Jaimes-Arroyo Rafael, Sánchez Olivia, Lara-Rojas Fernando, Aguirre Jesús
Departamento de Biología Celular y del Desarrollo, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City, Mexico.
Posgrado en Ciencias Biológicas, Unidad de Posgrado, Mexico City, Mexico.
Front Microbiol. 2018 Oct 23;9:2518. doi: 10.3389/fmicb.2018.02518. eCollection 2018.
Stress activated MAP kinases (SAPKs) of the Hog1/Sty1/p38 family are specialized in transducing stress signals. In contrast to what is seen in animal cells, very few fungal species contain more than one SAPK. and other Aspergilli contain two SAPKs called SakA/HogA and MpkC. We have shown that SakA is essential for conidia to maintain their viability and to survive high HO concentrations. HO induces SakA nuclear accumulation and its interaction with transcription factor AtfA. Although SakA and MpkC show physical interaction, little is known about MpkC functions. Here we show that mutants are not sensitive to oxidative stress but in fact MpkC inactivation partially restores the oxidative stress resistance of mutants. mutants display about twofold increase in the production of fully viable conidia. The inactivation of the SakA upstream MAPKK PbsB or the simultaneous elimination of and result in virtually identical phenotypes, including decreased radial growth, a drastic reduction of conidiation and a sharp, progressive loss of conidial viability. SakA and to a minor extent MpkC also regulate cell-wall integrity. Given the roles of MpkC in conidiation and oxidative stress sensitivity, we used a functional MpkC::GFP fusion to determine MpkC nuclear localization as an indicator of MpkC activation during asexual development and stress. MpkC is mostly localized in the cytoplasm of intact conidia, accumulates in nuclei during the first 2 h of germination and then becomes progressively excluded from nuclei in growing hyphae. In the conidiophore, MpkC nuclear accumulation increases in vesicles, metulae and phialides and decreases in older conidia. Oxidative and osmotic stresses induce MpkC nuclear accumulation in both germinating conidia and hyphae. In all these cases, MpkC nuclear accumulation is largely dependent on the MAPKK PbsB. Our results indicate that SakA and MpkC play major, distinct and sometimes opposing roles in conidiation and conidiospore physiology, as well as common roles in response to stress. We propose that two SAPKs are necessary to delay (MpkC) or fully stop (SakA) mitosis during conidiogenesis and the terminal differentiation of conidia, in the highly prolific phialoconidiation process characteristic of the Aspergilli.
Hog1/Sty1/p38家族的应激激活丝裂原活化蛋白激酶(SAPK)专门负责转导应激信号。与动物细胞不同,很少有真菌物种含有不止一种SAPK。烟曲霉和其他曲霉含有两种SAPK,即SakA/HogA和MpkC。我们已经表明,SakA对于分生孢子维持其活力和在高浓度过氧化氢(HO)下存活至关重要。HO诱导SakA核积累及其与转录因子AtfA的相互作用。尽管SakA和MpkC显示出物理相互作用,但对MpkC的功能了解甚少。在这里我们表明,(此处原文缺失相关突变体信息)突变体对氧化应激不敏感,但实际上MpkC失活部分恢复了(此处原文缺失相关突变体信息)突变体的氧化应激抗性。(此处原文缺失相关突变体信息)突变体在完全可存活分生孢子的产生上显示出约两倍的增加。SakA上游的丝裂原活化蛋白激酶激酶(MAPKK)PbsB失活或同时消除(此处原文缺失相关信息)和(此处原文缺失相关信息)会导致几乎相同的表型,包括径向生长减少、分生孢子形成急剧减少以及分生孢子活力急剧、渐进性丧失。SakA以及在较小程度上MpkC也调节细胞壁完整性。鉴于MpkC在分生孢子形成和氧化应激敏感性方面的作用,我们使用功能性MpkC::GFP融合蛋白来确定MpkC的核定位,作为无性发育和应激过程中MpkC活化的指标。MpkC大多定位于完整分生孢子的细胞质中,在萌发的最初2小时内在细胞核中积累,然后在生长的菌丝中逐渐从细胞核中排除。在分生孢子梗中,MpkC在小泡、梗基和产孢细胞中的核积累增加,而在较老的分生孢子中减少。氧化应激和渗透应激在萌发的分生孢子和菌丝中均诱导MpkC核积累。在所有这些情况下,MpkC核积累在很大程度上依赖于MAPKK PbsB。我们的结果表明,SakA和MpkC在分生孢子形成和分生孢子生理中发挥主要、不同且有时相反的作用,以及在应激反应中发挥共同作用。我们提出,在曲霉特有的高产瓶梗分生孢子形成过程中,两种SAPK对于在分生孢子发生和分生孢子的终末分化过程中延迟(MpkC)或完全停止(SakA)有丝分裂是必要的。