The guide sRNA sequence determines the activity level of box C/D RNPs.

Graziadei A, Gabel F, Kirkpatrick J, Carlomagno T
Elife 9 (2020 Mar 23)

PMID: 32202498 doi: 10.7554/eLife.50027

Submitted to SASBDB: 2019 Jul 29 Published in SASBDB: 2020 Mar 25

SASDGV2 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Nop5, 1H L7Ae, 1H st-sR26 RNA, 1H substrate D' and 2H Fibrillarin (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at KWS1, FRM2 on 2015 May 24 RgGuinier 4.9 nm Dmax 16.0 nm VolumePorod 87 nm3

SASDGW2 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H L7Ae, 1H st-sR26 RNA, 1H substrate D' and 2H Nop5 (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2015 Sep 21 RgGuinier 4.1 nm Dmax 14.0 nm VolumePorod 210 nm3

SASDGX2 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H Nop5, 1H st-sR26 RNA, 1H substrate D' and 2H L7Ae (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2015 Nov 23 RgGuinier 4.0 nm Dmax 13.3 nm VolumePorod 42 nm3

SASDGY2 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H Nop5, 1H L7Ae, 1H substrate D' and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2019 Jul 21 RgGuinier 2.8 nm Dmax 9.9 nm VolumePorod 52 nm3

SASDGZ2 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Nop5, 1H L7Ae, 1H substrate D', 2H Fibrillarin and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at KWS1, FRM2 on 2015 May 23 RgGuinier 4.8 nm Dmax 16.6 nm VolumePorod 82 nm3

SASDG23 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H L7Ae, 1H substrate D', 2H(70%) Nop5 and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2018 Apr 8 RgGuinier 4.0 nm Dmax 13.8 nm VolumePorod 214 nm3

SASDG33 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H L7Ae, 1H substrate D, 2H(70%) Nop5 and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2018 Apr 8 RgGuinier 4.0 nm Dmax 13.9 nm VolumePorod 231 nm3

SASDG43 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Nop5, 1H L7Ae, 1H st-sR26 RNA, 1H substrate D and 2H Fibrillarin (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at KWS1, FRM2 on 2015 May 24 RgGuinier 4.9 nm Dmax 16.6 nm VolumePorod 65 nm3

SASDG53 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Nop5, 1H L7Ae, 1H st-sR26 RNA, 1H substrate D and 2H Nop5 (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2015 Sep 21 RgGuinier 4.1 nm Dmax 14.0 nm VolumePorod 222 nm3

SASDG63 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H Nop5, 1H st-sR26 RNA, 1H substrate D and 2H L7Ae (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2015 Nov 23 RgGuinier 3.9 nm Dmax 12.5 nm VolumePorod 49 nm3

SASDG73 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Fibrillarin, 1H Nop5, 1H L7Ae, 1H substrate D and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at D22, Institut Laue-Langevin (ILL) on 2019 Jul 21 RgGuinier 2.7 nm Dmax 9.5 nm VolumePorod 25 nm3

SASDG83 – The Pyrococcus furiosus Box C/D sRNP complex reconstituted with 1H Nop5, 1H L7Ae, 1H substrate D, 2H Fibrillarin and 2H st-sR26 RNA (SANS data in 42% v/v D2O) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl 42%D2O, pH: 6.6 Experiment: SANS data collected at KWS1, FRM2 on 2015 May 23 RgGuinier 5.1 nm Dmax 15.5 nm VolumePorod 125 nm3

SASDG93 – Pyrococcus furiosus Box C/D sRNP reconstituted with st-sR6 sRNA bound to substrate D' (SAXS data) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D' monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl, pH: 6.6 Experiment: SAXS data collected at BM29, ESRF on 2015 Jun 13 RgGuinier 4.7 nm Dmax 15.8 nm VolumePorod 346 nm3

SASDGA3 – Pyrococcus furiosus Box C/D sRNP reconstituted with st-sR6 sRNA bound to substrate D (SAXS data) Sample: Fibrillarin-like rRNA/tRNA 2'-O-methyltransferase dimer, 52 kDa Pyrococcus furiosus protein 50S ribosomal protein L7Ae dimer, 27 kDa Pyrococcus furiosus protein NOP5/NOP56 related protein dimer, 94 kDa Pyrococcus furiosus protein Pyrococcus furiosus sR26 stabilized construct monomer, 24 kDa Pyrococcus furiosus RNA Pyrococcus furiosus sR26 substrate D monomer, 4 kDa Pyrococcus furiosus RNA Buffer: 50 mM phosphate 500 mM NaCl, pH: 6.6 Experiment: SAXS data collected at BM29, ESRF on 2015 Jul 13 RgGuinier 5.0 nm Dmax 17.0 nm VolumePorod 429 nm3