Data from: Brucella NyxA and NyxB dimerization enhances effector function during infection
Data files
May 20, 2025 version files 739.56 KB
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RawData.pdf
737.18 KB
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README.md
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Abstract
Supporting data (uncropped RAW western blots) for the article entitled: Brucella NyxA and NyxB dimerization enhances effector function during infection
Data concern two Brucella translocated effectors, NyxA and NyxB, that contribute to the late stages of intracellular multiplication. We show that 1) NyxA and NyxB directly interact; 2) monomeric forms of the Nyx effectors still interact with their host cellular target, the deSUMOylase sentrin-specific protease 3 (SENP3); 3) monomers are less able to delocalize SENP3 from the nucleoli.
Dataset DOI: 10.5061/dryad.fxpnvx14s
Description of the data and file structure
The dataset presented corresponds to the uncropped gels and western blots used in the manuscript entitled: Study of the molecular interactions underlying the Brucella NyxA and NyxB effector function.
File: RawData.pdf
Figure 1. In vitro and in cellullo interaction between NyxA and NyxB.
(A) Pulldown using purified NyxB against His-NyxA immobilised on a Ni NTA resin or the inverse (B) His-NyxB immobilized. For both, an empty column was used as a control for non-specific binding. Interactions were visualised with coomassie blue stained gels. The flowthrough (FT), wash (W) and elution (E) fractions are shown for each sample. (D) Co-immunoprecipitation (co-IP) assay from cells expressing NyxA or NyxB tagged with HA or Myc using anti-HA-magnetic beads. The input and co-IP fractions were revealed using an anti-Myc, and an anti-HA antibodies. Molecular weights are indicated (kDa).
Figure 3. Monomeric NyxA and NyxB still target the nucleus and are part of the same complex.
(B) Co-immunoprecipitation (co-IP) assay from cells expressing NyxA-L93E or NyxB-L97E tagged with HA or Myc. The input and co-IP fractions were revealed using an anti-Myc, an anti-HA antibody and anti-Nyx antibody. Molecular weights are indicated (kDa).
Figure 4. Monomeric NyxA and NyxB still interact with SENP3.
(A) Co-immunoprecipitation (co-IP) assay suing GFP-Trap, incubated with lysates from cells expressing GFP-SENP3 and Myc-NyxA, NyxB, NyxA-L93E or NyxB-L97E. GFP was used as a control for nonspecific binding. The inputs and co-IP elutions were revealed using an anti-Myc antibody, and an anti-GFP antibody. Molecular weights are indicated (kDa). (B) Co-immunoprecipitation (co-IP) assay using anti-HA magnetic bead, incubated with lysates from cells expressing HA-NyxA, HA-NyxA-L93E, HA-NyxB or HA-NyxB-L97E. Cells transfected with the HA empty vector were used as a control for nonspecific binding. The inputs and co-IP elutions were revealed using an anti-HA antibody, and an anti-SENP3 antibody. Molecular weights are indicated (kDa).
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