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1.
Figure 4

Figure 4. Copper-dependent standard and alternative respiration in P. anserina.. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

While respiration of the wild type predominantly proceeds via the standard copper-dependent pathway, respiration in the grisea mutants does mainly follow the alternative route utilizing the di-iron containing alternative oxidase.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.
2.
Figure 6

Figure 6. Transcript analysis of Pa_2_7880 (PaMth1: A, B) and Pa_2_7310 (C, D).. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

The results of the SuperSAGE analysis (A, C) are shown as tags per million (tpm) for wild type (WT) and grisea (gr). The results of the qRT-PCR (B, D) are relative expression levels of the gene of the wild type (WT, n = 6) and of grisea (gr, n = 6) grown in standard medium, and wild type (WT+Cu, n = 3) and grisea (gr+Cu, n = 3) grown on copper supplemented medium, normalized to the expression level of the gene coding for mitochondrial PORIN. The error bars represent the standard deviation.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.
3.
Figure 2

Figure 2. High and low affinity copper uptake of copper and delivery to different enzymes and compartments.. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

At cellular copper levels, copper-uptake is known from S. cerevisiae and P. anserina to involve high affinity uptake mechanism. After reduction of copper mediated by a membrane located oxidoreductase (FRE) the import is performed by high affinity copper transporters (CTR). Expression of CTR-transporter genes is regulated transcriptionally by copper-dependent transcription factors. Intracellularly the copper is stored and distributed via copper binding chaperones and partially becomes transported into mitochondrion where it is stored and distributed to copper-containing proteins (i.e. cytochrome oxidase). Copper is also taken up by cells via a low copper-uptake system.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.
4.
Figure 1

Figure 1. Transcript analysis of Pa_1_16400 (PaCtr1: A, B), Pa_4_4770 (PaCtr2: C, D), and Pa_3_10440 (PaCtr3: E, F), Pa_5_11970 (G, H), Pa_3_1710 (I, J).. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

Results from the SuperSAGE analysis (A, C, E, G, I) are shown as ‘tags per million’ (tpm) for wild type (WT) and grisea (gr). The results of the qRT-PCR (B, D, F, H, J) are relative expression levels of the three copper transporter genes of the wild type (WT, n = 5 in B, D, F, J; n = 6 in H) and the grisea mutant (gr, n = 6) grown in standard medium, and wild type (WT+Cu, n = 3) and grisea (gr+Cu, n = 3) grown on copper-supplemented medium, normalized to the expression level of the gene coding for mitochondrial PORIN The error bars represent the standard deviation.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.
5.
Figure 3

Figure 3. High and low affinity uptake of iron in filamentous fungi.. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

Under iron limiting conditions two high affinity iron import mechanisms are involved in uptake of iron from the environment. Via the reductive iron assimilation (RIA) system, ferric iron first becomes reduced to ferrous iron by a reductase (FRE) and then oxidized by a multicopper oxidase (FET) and imported by a plasma membrane transporter (FTR). Many fungi also produce and secrete siderophores (SID) and, after binding of iron (SID Fe) are reimported into the cell. Bound iron is subsequently made accessible to the cell by activity of a specific esterase (ESTB). In S. cerevisiae is only able to take up siderophores (Xenosiderophores) that available in the environment.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.
6.
Figure 5

Figure 5. Transcript (A, B) and protein analysis (C) of Pa_2_4660 (PaSod2).. From: A Differential Genome-Wide Transcriptome Analysis: Impact of Cellular Copper on Complex Biological Processes like Aging and Development.

The results of the SuperSAGE analysis (A) are shown as tags per million (tpm) for wild type (WT) and grisea (gr). The results of the qRT-PCR (B) are relative expression levels of the gene of the wild type (WT, n = 5) and of grisea (gr, n = 6) grown in standard medium, and wild type (WT+Cu, n = 3) and grisea (gr+Cu, n = 3) grown on copper supplemented medium, normalized to the expression level of gene coding for mitochondrial PORIN. The error bars represent the standard deviation. For Western Blot (α-PaSOD2) and in-gel SOD activity assay (C) 100 µg of total protein extracts of three independent wild type (WT) and grisea (gr) isolates were used. As loading control the α-PaPRE3 antibody was used.

Jörg Servos, et al. PLoS One. 2012;7(11):e49292.

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