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. 2025 Jul 21;16(8):e00845-25. doi: 10.1128/mbio.00845-25

Fig 11.

ChCpc1-ChChk1 protein interaction in Cochliobolus heterostrophus shown by two assays. Gene expression reveals that ChChk1 regulates autophagy and arginine metabolism, with ChARG and ChATG genes downregulated in mutants under MSX stress.

ChCpc1 interacted with ChChk1 in Cochliobolus heterostrophus. (A) ChCpc1 interacted with ChChk1 in yeast two-hybrid assay. Serial dilutions of the yeast cells were plated on synthetic dropout (SD) medium that lacked histidine (H), leucine (L), and tryptophan (T) (SD- T/H/L). The interaction between pGADT7-T and ChChk1 was used as control. (B) Pull-down assay to verify the interaction between ChCpc1 and ChChk1. The fusion protein His-Cpc1 was detected in the eluted solution of GST-ChChk1 and His-ChCpc1 co-incubated mixture but not in that of glutathione S-transferase (GST) and His-ChCpc1. (C) Expression levels of ChARG1, ChARG4, ChATG5, ChATG7, ChATG8, and ChATG9 in WT and Δchk1 mutants under different culture conditions. CM: complete medium; MSX: L-methionine-DL-sulfoximine; MM: minimal medium; MM + MSX: minimal medium supplemented with 100 µM L-methionine-DL-sulfoximine (MSX). GraphPad Prism program’s t-tests and multiple t-tests were used for the analysis of significant differences. The bars indicate the standard error of means of three replications.