CRYSTAL STRUCTURE OF SRP19 IN COMPLEX WITH THE S DOMAIN OF SIGNAL RECOGNITION PARTICLE RNA

CYTIDINE-5'-PHOSPHATE-2',3'-CYCLIC PHOSPHATE

Crystal structure of SRP19 in complex with the S domain of SRP RNA and its implication for the assembly of the signal recognition particle.

MUTANT (C112G, G113A, G238U, G239C) 7SL RNA (NUCLEOTIDES 112-239)
signal recognition particle RNA S domain

SIGNAL RECOGNITION PARTICLE 19 KDA PROTEIN

GACACUAAGUUCGGCAUCAAUAUGGUGACCUCCCGGGAGCGGGGGACCACCAGGUUGCCUAAGGAGGGGUGAACCGGCCC
AGGUCGGAAACGGAGCAGGUCAAAACUCCCGUGCUGAUCAGUAGUGU(CCC)

GACACUAAGUUCGGCAUCAAUAUGGUGACCUCCCGGGAGCGGGGGACCACCAGGUUGCCUAAGGAGGGGUGAACCGGCCC
AGGUCGGAAACGGAGCAGGUCAAAACUCCCGUGCUGAUCAGUAGUGUC

MIIWPSYIDKKKSRREGRKVPEELAIEKPSLKDIEKALKKLGLEPKIYRDKRYPRQHWEIAGRVEVDYKGNKLCLLKEIA
KIIKGKN

ORIGINAL PHASES WERE DETERMINED BY SELENOMETHIONINE MAD BUT FINAL REFINEMENT WAS CARRIED OUT AGAINST A HIGHER QUALITY SINGLE WAVELENGTH DATASET FOR WHICH WE SUPPLY STATISTICS.

12% PEG4000, 110MM Tris.HCl pH 7.4, 10% glycerol, 50mm magnesium acetate, 100MM ammonium acetate, 5MM magnesium chloride, VAPOR DIFFUSION, HANGING DROP, temperature 300.5K

This sequence is 7SL RNA from Homo sapiens. 128 nt RNA fragment was in vitro transcribed under control of a T7 promoter with T7 RNA polymerase. It was Cotranscribed with 2 cis-acting hammerhead ribozymes to cleave homogeneous RNA from the full length transcript.

Refmac5.1 (Murshudov) was also used in refinement.

PROTEIN-RNA COMPLEX, RIBONUCLEOPROTEIN, SRP, SIGNAL RECOGNITION PARTICLE, TETRALOOP, SIGNALING PROTEIN-RNA COMPLEX

MIIWPSYIDKKKSRREGRKVPEELAIEKPSLKDIEKALKKLGLEPKIYRDKRYPRQHWEICGCVEVDYKGNKLQLLKEIC
KIIKGKN

CGCACTAAGTTCGGCATCAATATGGTGACCTCCCGGGAGCGGGGGACCACCAGGTTGCCTAAGGAGGGGTGAACCGGCCC
AGGTCGGAAACGGAGCAGGTCAAAACTCCCGTGCTGATCAGTAGTGGG