TY - JOUR

T1 - Velvet domain protein VosA represses the zinc cluster transcription factor ScIB regulatory network for Aspergillus nidulans asexual development, oxidative stress response and secondary metabolism

AU - Thieme, Karl G.

AU - Gerke, Jennifer

AU - Sasse, Christoph

AU - Valerius, Oliver

AU - Thieme, Sabine

AU - Karimi, Razieh

AU - Heinrich, Antje K.

AU - Finkernagel, Florian

AU - Smith, Kristina

AU - Bode, Helge B.

AU - Freitag, Michael

AU - Ram, Arthur F. J.

AU - Braus, Gerhard H.

N1 - Funding information:Thispaperwassupportedbythe followinggrants:Deutsche Forschungsgemeinschaft(DFG):BR1502/11-2 andSFB860.HBBacknowledgestheDeutsche ForschungsgemeinschaftforfundingoftheImpact IIqTofmassspectrometer(INST161/810-1).The

PY - 2018/7/25

Y1 - 2018/7/25

N2 - There is an error in panel D of S8 Fig. Specifically, the upper panel should read ‘SclB-cYFP + nYFP’, not ‘SclA-cYFP + nYFP’. The authors have provided a corrected version here. Supporting information S8 Fig GFP-fusion proteins of SclB are functional and phosphorylated and Bi-FC controls are negative. A) Strains expressing SclB either N- or C-terminally tagged with sGFP in ΔsclB background, ΔsclB and wildtype (WT) were point inoculated on solid MM and grown for 4 days in light. B) SclB-GFP and GFP-SclB fusion proteins expressed under native promoter are visualized in a western hybridization assay employing an α-GFP antibody (GFP) and Ponceau staining as loading control (Pnc). The black arrow indicates bands corresponding to full-length fusion proteins (in silico prediction 87.46 kDa). C) Protein crude extracts of GFP-SclB grown vegetatively were mixed with phosphatase inhibitor cocktail (-/PhoI), with Lambda phosphatase (λ/-), or Lambda phosphatase and phosphatase inhibitor cocktail (λ/PhoI). A control sample was left untreated (-/-). A subsequent western hybridization assay employing α-GFP antibody visualizes protein bands. D) Two strains, either expressing sclB::cyfp and the free second half of the split YFP (nyfp; upper part), or free cyfp and rcoA::nyfp (lower part), under control of a bi-directional nitrate promoter were constructed. Strains were inoculated in liquid MM and analyzed with fluorescence microscopy after 36 h at 30°C. (TIF).

AB - There is an error in panel D of S8 Fig. Specifically, the upper panel should read ‘SclB-cYFP + nYFP’, not ‘SclA-cYFP + nYFP’. The authors have provided a corrected version here. Supporting information S8 Fig GFP-fusion proteins of SclB are functional and phosphorylated and Bi-FC controls are negative. A) Strains expressing SclB either N- or C-terminally tagged with sGFP in ΔsclB background, ΔsclB and wildtype (WT) were point inoculated on solid MM and grown for 4 days in light. B) SclB-GFP and GFP-SclB fusion proteins expressed under native promoter are visualized in a western hybridization assay employing an α-GFP antibody (GFP) and Ponceau staining as loading control (Pnc). The black arrow indicates bands corresponding to full-length fusion proteins (in silico prediction 87.46 kDa). C) Protein crude extracts of GFP-SclB grown vegetatively were mixed with phosphatase inhibitor cocktail (-/PhoI), with Lambda phosphatase (λ/-), or Lambda phosphatase and phosphatase inhibitor cocktail (λ/PhoI). A control sample was left untreated (-/-). A subsequent western hybridization assay employing α-GFP antibody visualizes protein bands. D) Two strains, either expressing sclB::cyfp and the free second half of the split YFP (nyfp; upper part), or free cyfp and rcoA::nyfp (lower part), under control of a bi-directional nitrate promoter were constructed. Strains were inoculated in liquid MM and analyzed with fluorescence microscopy after 36 h at 30°C. (TIF).

UR - http://www.scopus.com/inward/record.url?scp=85050995872&partnerID=8YFLogxK

U2 - 10.1371/journal.pgen.1007511

DO - 10.1371/journal.pgen.1007511

M3 - Article

VL - 14

JO - PLoS Genetics

JF - PLoS Genetics

SN - 1553-7404

IS - 7

M1 - e1007511

ER -