Contrasting DNA-binding behaviour by ISL1 and LHX3 underpins differential gene targeting in neuronal cell specification

Smith N, Wilkinson-White L, Kwan A, Trewhella J, Matthews J
Journal of Structural Biology: X :100043 (2020 Dec)

doi: 10.1016/j.yjsbx.2020.100043

Submitted to SASBDB: 2020 Oct 1 Published in SASBDB: 2020 Dec 20

SASDJB9 – LIM/homeobox protein Lhx3 homeodomain (Lhx3-HD): HD3 Sample: LIM/homeobox protein Lhx3 monomer, 10 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 2.1 nm Dmax 7.0 nm VolumePorod 18 nm3

SASDJC9 – Insulin gene enhancer protein Isl-1 LIM-interaction domain (Isl1-LID) fused to the LIM/homeobox protein Lhx3 LIM-homeodomain (Lhx3-LIMHD): LLHD3 Sample: LIM/homeobox protein Lhx3 monomer, 23 kDa Mus musculus protein Insulin gene enhancer protein ISL-1 monomer, 4 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 3.3 nm Dmax 12.0 nm VolumePorod 39 nm3

SASDJD9 – Insulin gene enhancer protein Isl-1 homeodomain (Isl1-HD) fused to the LIM/homeobox protein Lhx3 homeodomain (Lhx3-HD): 2HD Sample: Insulin gene enhancer protein ISL-1 monomer, 12 kDa Mus musculus protein LIM/homeobox protein Lhx3 monomer, 9 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 3.4 nm Dmax 12.7 nm VolumePorod 40 nm3

SASDJE9 – Insulin gene enhancer protein Isl-1 homeodomain/LIM-interaction domain (Isl1-HDLID) fused to the LIM/homeobox protein Lhx3 LIM-homeodomain (Lhx3-LIMHD): 2HDLL Sample: Insulin gene enhancer protein ISL-1 monomer, 14 kDa Mus musculus protein LIM/homeobox protein Lhx3 monomer, 23 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 3.4 nm Dmax 13.0 nm VolumePorod 76 nm3

SASDJF9 – M100 DNA oligonucleotide bound to the LIM/homeobox protein Lhx3 homeodomain (Lhx3-HD): M100 + HD3 Sample: M100 oligonucleotide monomer, 12 kDa DNA LIM/homeobox protein Lhx3 monomer, 10 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 2.1 nm Dmax 7.0 nm VolumePorod 27 nm3

SASDJG9 – M100 DNA oligonucleotide bound to the protein fusion of Insulin gene enhancer protein Isl-1 LIM-interaction domain (Isl1-LID) and LIM/homeobox protein Lhx3 LIM-homeodomain (Lhx3-LIMHD): M100 + LLHD3 Sample: M100 oligonucleotide monomer, 12 kDa DNA LIM/homeobox protein Lhx3 monomer, 23 kDa Mus musculus protein Insulin gene enhancer protein ISL-1 monomer, 4 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 3.6 nm Dmax 13.0 nm VolumePorod 49 nm3

SASDJH9 – M100 DNA oligonucleotide bound to the protein fusion of Insulin gene enhancer protein Isl-1 homeodomain (Isl1-HD) and LIM/homeobox protein Lhx3 homeodomain (Lhx3-HD): M100 + 2HD Sample: M100 oligonucleotide monomer, 12 kDa DNA Insulin gene enhancer protein ISL-1 monomer, 12 kDa Mus musculus protein LIM/homeobox protein Lhx3 monomer, 9 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 2.4 nm Dmax 7.5 nm VolumePorod 41 nm3

SASDJJ9 – M100 DNA oligonucleotide bound to the protein fusion of Insulin gene enhancer protein Isl-1 homeodomain/LIM-interaction domain (Isl1-HDLID) and LIM/homeobox protein Lhx3 LIM-homeodomain (Lhx3-LIMHD): M100 + 2HDLL Sample: M100 oligonucleotide monomer, 12 kDa DNA Insulin gene enhancer protein ISL-1 monomer, 14 kDa Mus musculus protein LIM/homeobox protein Lhx3 monomer, 23 kDa Mus musculus protein Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 3.6 nm Dmax 14.0 nm VolumePorod 70 nm3

SASDJK9 – M100 DNA oligonucleotide Sample: M100 oligonucleotide monomer, 12 kDa DNA Buffer: 20 mM sodium phosphate monobasic/dibasic, 100 mM NaCl, 1 mM DTT, pH: 7.4 Experiment: SAXS data collected at SAXS/WAXS, Australian Synchrotron on 2018 Nov 2 RgGuinier 1.9 nm Dmax 6.8 nm VolumePorod 15 nm3