Alternative dimerization is required for activity and inhibition of the HEPN ribonuclease RnlA

Garcia-Rodriguez G, Charlier D, Wilmaerts D, Michiels J, Loris R
Nucleic Acids Research 49(12):7164-7178 (2021 Jun 17)

doi: 10.1093/nar/gkab513

Submitted to SASBDB: 2020 Mar 5 Published in SASBDB: 2021 May 31

SASDHW7 – RnlA-RnlB toxin-antitoxin complex (mRNA endoribonuclease toxin LS : Antitoxin RnlB) Sample: MRNA endoribonuclease toxin LS dimer, 80 kDa Escherichia coli protein Antitoxin RnlB (C-terminal His-tagged) monomer, 15 kDa Escherichia coli protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, 5% glycerol, pH: 8 Experiment: SAXS data collected at BM29, ESRF on 2018 Jul 25 RgGuinier 3.5 nm Dmax 10.0 nm VolumePorod 161 nm3

SASDHX7 – Escherichia coli RnlA (mRNA endoribonuclease toxin LS) Sample: MRNA endoribonuclease toxin LS dimer, 80 kDa Escherichia coli protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, 5% glycerol, pH: 8 Experiment: SAXS data collected at BM29, ESRF on 2018 Jul 25 RgGuinier 3.7 nm Dmax 10.7 nm VolumePorod 134 nm3

SASDKL2 – NRD-HEPN - mRNA endoribonuclease toxin LS from Escherichia coli (strain K12) Sample: NRD-HEPN truncated variant of RnlA endoribonuclease dimer, 62 kDa Escherichia coli protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, pH: 8 Experiment: SAXS data collected at SWING, SOLEIL on 2020 Jun 27 RgGuinier 3.0 nm Dmax 10.4 nm VolumePorod 88 nm3

SASDHY7 – Escherichia coli RnlA (mRNA endoribonuclease toxin LS) - R255A single alanine mutant Sample: MRNA endoribonuclease toxin LS - R255A mutant; N-terminal His-tagged dimer, 84 kDa Escherichia coli protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, 5% glycerol, pH: 8 Experiment: SAXS data collected at BM29, ESRF on 2018 Jul 25 RgGuinier 3.7 nm Dmax 20.1 nm VolumePorod 133 nm3

SASDKS9 – mRNA endoribonuclease toxin LS (D245R mutant); batch-SAXS measurements in low salt buffer Sample: MRNA endoribonuclease toxin LS (D245R mutant) dimer, 84 kDa Escherichia coli (strain … protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, pH: 8 Experiment: SAXS data collected at SWING, SOLEIL on 2020 Jul 18 RgGuinier 4.3 nm Dmax 15.9 nm VolumePorod 183 nm3

SASDHZ7 – Escherichia coli RnlA (mRNA endoribonuclease toxin LS) - R318A single alanine mutant Sample: MRNA endoribonuclease toxin LS - R318A mutant; N-terminal His-tagged dimer, 84 kDa Escherichia coli protein Buffer: 20 mM Tris, 150 mM NaCl, 1 mM TCEP, 5% glycerol, pH: 8 Experiment: SAXS data collected at BM29, ESRF on 2018 Jul 25 RgGuinier 3.4 nm Dmax 12.7 nm VolumePorod 124 nm3