Table 1.
Neurospora crassa strains used in this study.
Figure 1.
Changes in arginine methylation profiles in PRMT-encoding gene deletion strains.
Western-blot analysis of total cell extracts probed with: (A) SYM24 antibodies used to identify asymmetric dimethylarginine and (B) SYM10 antibodies used to identify symmetric dimethylarginine. The most prominent proteins in which differences in the Arg-methylated signals were detected are marked by arrows.
Figure 2.
Effect of Polyoxin D and Voriconazole on growth of N. crassa PRMT mutants.
All strains were cultured on sorbose (2%)-amended (which restricts colony growth) medium at 34°C. (A) control (B) with PolyoxinD (6.5 mM) (C) Voriconazole (12.23 mM).
Table 2.
Morphological consequences of deletion in PRMT-encoding genes.
Figure 3.
Co- Immunoprecipitation of MYC-SKB1 and COT1.
Corresponding proteins were detected with anti-Myc, in wild-type and cot-1 ts backgrounds. The experiment was performed under restrictive temperature (36°C). “|Cell extract” represents control without immunoprecipitation.
Figure 4.
Deletion of skb-1 affects COT1 protein level.
Western-blot analysis of total cell extracts probed with anti-Myc antibodies (A) Single and double PRMT deletion strains in a MYC::COT1 background (B) MYC::COT1 in aΔskb-1 background at two developmental stages. β tubulin levels are presented as controls for each developmental stage examined.. MYC::COT1 isoforms are marked by arrows.
Figure 5.
Deletion of skb-1 does not affect COT1 interaction with MOB2 co-activators.
Immunoprecipitation of MYC::COT1 with anti-Myc antibodies. Proteins obtained were resolved on a gradient gel and subjected to silver staining. MOB2A and MOB2B co-precipitants are marked. MYC::COT1 isoforms are marked by arrows. * marks the IgG.
Figure 6.
Arginine methylation on MYC::COT1.
Immunoprecipitation of MYC::COT1with anti-Myc antibodies probed with: (A) SYM24 antibodies (detecting asymmetric dimethylarginine) and (B) SYM10 antibodies (detecting symmetric dimethylarginine). MYC::COT1 isoforms are marked by arrows.
Figure 7.
Colony morphology of PRMT- encoding gene deletion strains in a cot-1 ts background.
Strains were cultured for 72 hours at 30°C (a semi-restrictive temperature).
Table 3.
Growth rate of PRMT-encoding genes deletion strains in a cot-1 ts background.
Figure 8.
Effect of inactivation of Δamt-3 or Δskb-1 on growth of cot-1 ts.
cot-1 ts, grown in the presence of NaCl, exhibits a suppressed colonial phenotype. The colonial phenotype is further suppressed upon inactivation of amt-3 or skb-1. Strains were cultured for 72 hours at 34°C (cot-1 ts restrictive temperature) with 0.5, 0.75, 1 or 1.25 M NaCl. Colony area is presented as percent of control. * and + indicate a significant difference (p<0.05) of colony area in the Δamt-3;cot-1 or Δskb-1;cot-1 backgrounds, respectively.