Figure 4. S. marcescens chitinases are... | Rockefeller University Press

Figure 4.

$Figure 4. S. marcescens chitinases are secreted in chiW- and chiX-dependent manner. (A) S. marcescens (Db10) together with JJH04w (ΔchiW), JJH05x (ΔchiX), JJH06y (ΔchiY), and JJH07z (ΔchiZ) were analyzed by Western immunoblotting using the specific antisera indicated. (B) Complementation of the ΔchiW strain. The JJH04w strain was transformed with pUNI-PROM or pUNI-ChiW (pChiW), and the localization of ChiC was analyzed. (C) Complementation of the ΔchiX strain. The JJH05x strain was transformed with pUNI-PROM or pUNI-ChiX (pChiX), and the localization of ChiC was analyzed. (D) S. marcescens (Db10) together with the JJH04w (ΔchiW) mutant and the JJH05x (ΔchiX) mutant were fractionated into cytoplasm (C), periplasm (P), total membranes (M), and culture supernatant (SN). Proteins were analyzed by Western immunoblotting using the antisera indicated. Controls were the periplasmic maltose binding protein (MBP) and the cytoplasmic glutamine synthetase (GS) enzyme. In all cases, the band marked by the single asterisk is the ChiA protein, which the polyclonal ChiB antiserum can also detect. (E) The FTG005 strain (ϕchiA::gfp) and the FTG006 strain (ϕchiA::gfp and ΔchiX::mKate) were grown in rich media before the localization of ChiC and GFP was analyzed by Western immunoblotting. WC, whole cell.$

S. marcescens chitinases are secreted in chiW- and chiX-dependent manner. (A) S. marcescens (Db10) together with JJH04w (ΔchiW), JJH05x (ΔchiX), JJH06y (ΔchiY), and JJH07z (ΔchiZ) were analyzed by Western immunoblotting using the specific antisera indicated. (B) Complementation of the ΔchiW strain. The JJH04w strain was transformed with pUNI-PROM or pUNI-ChiW (pChiW), and the localization of ChiC was analyzed. (C) Complementation of the ΔchiX strain. The JJH05x strain was transformed with pUNI-PROM or pUNI-ChiX (pChiX), and the localization of ChiC was analyzed. (D) S. marcescens (Db10) together with the JJH04w (ΔchiW) mutant and the JJH05x (ΔchiX) mutant were fractionated into cytoplasm (C), periplasm (P), total membranes (M), and culture supernatant (SN). Proteins were analyzed by Western immunoblotting using the antisera indicated. Controls were the periplasmic maltose binding protein (MBP) and the cytoplasmic glutamine synthetase (GS) enzyme. In all cases, the band marked by the single asterisk is the ChiA protein, which the polyclonal ChiB antiserum can also detect. (E) The FTG005 strain (ϕchiA::gfp) and the FTG006 strain (ϕchiA::gfp and ΔchiX::mKate) were grown in rich media before the localization of ChiC and GFP was analyzed by Western immunoblotting. WC, whole cell.