Soid-Raggi Gabriela, Sánchez Olivia, Aguirre Jesús
Departamento de Genética Molecular, Instituto de Fisiología Celular-UNAM, Apartado Postal 70-242, 04510, México, D.F., México.
Mol Microbiol. 2006 Feb;59(3):854-69. doi: 10.1111/j.1365-2958.2005.04996.x.
Asexual reproduction (conidiation) in Aspergillus nidulans is induced by environmental signals like exposure to air or nutrient starvation, and depends on brlA gene activation. The study of 'fluffy' mutants showing delayed asexual development and reduced brlA expression has defined the fluG pathway, involved in regulation of this differentiation process. Genetic characterization of a 'fluffy' mutant identified tmpA as a new gene involved in regulation of conidiation. TmpA defines a new family of putative transmembrane proteins of unknown function, widespread in filamentous fungi and plants, with homologues showing similarity to non-ribosomal peptide synthetases. The deletion of tmpA resulted in decreased brlA expression and conidiation in air-exposed colonies. This defect was suppressed when DeltatmpA mutants were grown next to wild-type or DeltafluG mutant colonies, even without direct contact between hyphae. In liquid culture, tmpA was essential for conidiation induced by nitrogen but not by carbon starvation, whereas the overexpression of different tmpA tagged alleles resulted in conidiation. The overexpression of fluG-induced conidiation independently of tmpA and DeltatmpADeltafluG double mutants showed an additive 'fluffy' phenotype, indicating that tmpA and fluG regulate asexual sporulation through different pathways. TmpA and its homologues appear to have diverged from the ferric reductase family, retaining overall transmembrane architecture, NAD(P), flavin adenine dinucleotide (FAD) and possibly haem-binding domains. Based on our results, we propose that TmpA is a membrane oxidoreductase involved in the synthesis of a developmental signal.
构巢曲霉中的无性繁殖(分生孢子形成)由诸如暴露于空气或营养饥饿等环境信号诱导,并依赖于brlA基因的激活。对表现出无性发育延迟和brlA表达降低的“蓬松”突变体的研究确定了fluG途径,该途径参与这一分化过程的调控。一个“蓬松”突变体的遗传特征鉴定表明tmpA是一个参与分生孢子形成调控的新基因。TmpA定义了一个功能未知的假定跨膜蛋白新家族,广泛存在于丝状真菌和植物中,其同源物与非核糖体肽合成酶具有相似性。tmpA的缺失导致暴露于空气中的菌落中brlA表达和分生孢子形成减少。当ΔtmpA突变体与野生型或ΔfluG突变体菌落相邻生长时,即使菌丝之间没有直接接触,这种缺陷也会受到抑制。在液体培养中,tmpA对于氮诱导而非碳饥饿诱导的分生孢子形成至关重要,而不同tmpA标签等位基因的过表达导致分生孢子形成。fluG诱导的分生孢子形成独立于tmpA,并且ΔtmpAΔfluG双突变体表现出累加的“蓬松”表型,表明tmpA和fluG通过不同途径调节无性孢子形成。TmpA及其同源物似乎已从铁还原酶家族分化而来,保留了整体跨膜结构、NAD(P)、黄素腺嘌呤二核苷酸(FAD)以及可能的血红素结合结构域。基于我们的结果,我们提出TmpA是一种参与发育信号合成的膜氧化还原酶。