Modulating the Cascade architecture of a minimal Type I-F CRISPR-Cas system.

Gleditzsch D, Müller-Esparza H, Pausch P, Sharma K, Dwarakanath S, Urlaub H, Bange G, Randau L
Nucleic Acids Res 44(12):5872-82 (2016 Jul 8)
PMID: 27216815
doi: 10.1093/nar/gkw469 		Submitted to SASBDB: 2016 Mar 18
Published in SASBDB:

SASDBK4 – The 1:1:3:1 crRNA:Cas6f:Cas7fv:Cas5fv CRISPR/Cas Type I-F short Cascade complex

short-crRNA: CRISPR/Cas Type I-F Cascade componentCas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family)Trimeric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821)Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822) experimental SAS data
DAMMIF model
Sample: Short-crRNA: CRISPR/Cas Type I-F Cascade component monomer, 14 kDa Shewanella putrefaciens RNA
Cas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family) monomer, 21 kDa Shewanella putrefaciens protein
Trimeric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821) trimer, 112 kDa Shewanella putrefaciens protein
Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822) monomer, 38 kDa Shewanella putrefaciens protein
Buffer: 50 mM HEPES 150 mM NaCl 1mM DTT 1mM EDTA, pH: 7
Experiment: SAXS data collected at BM29, ESRF on 2015 Jun 27
RgGuinier 4.1 nm
Dmax 14.2 nm

SASDBL4 – The 1:1:6:1 crRNA:Cas6f:Cas7fv:Cas5fv CRISPR/Cas Type I-F wild-type Cascade complex

Cas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family)Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822)Hexameric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821)wildtype-crRNA: CRISPR/Cas Type I-F Cascade component experimental SAS data
DAMMIF model
Sample: Cas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family) monomer, 21 kDa Shewanella putrefaciens protein
Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822) monomer, 38 kDa Shewanella putrefaciens protein
Hexameric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821) hexamer, 223 kDa Shewanella putrefaciens protein
Wildtype-crRNA: CRISPR/Cas Type I-F Cascade component monomer, 19 kDa RNA
Buffer: 50 mM HEPES 150 mM NaCl 1mM DTT 1mM EDTA, pH: 7
Experiment: SAXS data collected at BM29, ESRF on 2015 Jun 27
RgGuinier 5.4 nm
Dmax 18.4 nm

SASDBM4 – The 1:1:9:1 crRNA:Cas6f:Cas7fv:Cas5fv CRISPR/Cas Type I-F long Cascade complex

Cas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family)Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822)Nonameric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821)long-crRNA: CRISPR/Cas Type I-F Cascade component experimental SAS data
DAMMIF model
Sample: Cas6f: CRISPR/Cas Type I-F Cascade component (CRISPR-associated protein, Csy4 family) monomer, 21 kDa Shewanella putrefaciens protein
Cas5fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1822) monomer, 38 kDa Shewanella putrefaciens protein
Nonameric Cas7fv: CRISPR/Cas Type I-F Cascade component (Uncharacterized protein, Sputcn32_1821) nonamer, 335 kDa Shewanella putrefaciens protein
Long-crRNA: CRISPR/Cas Type I-F Cascade component monomer, 25 kDa RNA
Buffer: 50 mM HEPES 150 mM NaCl 1mM DTT 1mM EDTA, pH: 7
Experiment: SAXS data collected at BM29, ESRF on 2016 Jan 30
RgGuinier 6.5 nm
Dmax 21.6 nm