Manubens Augusto, Avila Marcela, Canessa Paulo, Vicuña Rafael
Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Casilla 114-D, Santiago, Chile.
Curr Genet. 2003 Sep;43(6):433-8. doi: 10.1007/s00294-003-0410-7. Epub 2003 Jun 11.
We previously identified and characterized three mnp genes coding for manganese peroxidase (MnP) in the white rot fungus Ceriporiopsis subvermispora. In this work, we assessed transcript levels of mnp genes in liquid cultures of this fungus grown under various conditions. In the absence of Mn(2+), mnp1 and mnp2 mRNA were detected by Northern hybridization, irrespective of the lack of extracellular MnP activity. Addition of Mn(2+) to the cultures led to a marked increase in both transcripts, the highest titers being observed at 10 micro M Mn(2+). mnp1 mRNA was not detected at Mn(2+ )concentrations above 80 micro M, whereas mnp2 mRNA was still observed at 320 micro M Mn(2+). Differential regulation of these genes was confirmed by the addition of Cu(2+), Zn(2+), Ag(+) and Cd(2+). These metal ions dramatically elevated both transcripts and also allowed the detection of the mnp3 transcript. In most cases, the increase in mRNA levels was partially abolished by the simultaneous presence of Mn(2+), although the latter was strictly required to detect extracellular MnP activity. However, the lignin-related compound syringic acid specifically increased the mnp1 transcript, although only in the absence of Mn(2+). These results indicate that there is no clear correlation between mnp mRNA levels and MnP activity. In addition, they strongly suggest that Mn(2+) plays a post-transcriptional role which is essential for the presence of active MnP in the extracellular fluid.
我们之前在白腐菌黄孢原毛平革菌中鉴定并表征了三个编码锰过氧化物酶(MnP)的mnp基因。在这项工作中,我们评估了该真菌在各种条件下进行液体培养时mnp基因的转录水平。在没有Mn(2+)的情况下,通过Northern杂交检测到了mnp1和mnp2 mRNA,尽管缺乏细胞外MnP活性。向培养物中添加Mn(2+)导致两种转录本显著增加,在10微摩尔Mn(2+)时观察到最高滴度。在Mn(2+)浓度高于80微摩尔时未检测到mnp1 mRNA,而在320微摩尔Mn(2+)时仍可观察到mnp2 mRNA。添加Cu(2+)、Zn(2+)、Ag(+)和Cd(2+)证实了这些基因的差异调节。这些金属离子显著提高了两种转录本水平,还使得能够检测到mnp3转录本。在大多数情况下,Mn(2+)的同时存在会部分消除mRNA水平的增加,尽管检测细胞外MnP活性严格需要Mn(2+)。然而,木质素相关化合物丁香酸特异性地增加了mnp1转录本,尽管仅在没有Mn(2+)的情况下。这些结果表明mnp mRNA水平与MnP活性之间没有明显的相关性。此外,它们强烈表明Mn(2+)发挥转录后作用,这对于细胞外液中活性MnP的存在至关重要。