Use of chemically modified nucleotides to determine a 62-nucleotide RNA crystal structure: a survey of phosphorothioates, Br, Pt and Hg.
Two important challenges confronting RNA crystallographers are producing crystals and finding isomorphous heavy-atom derivatives. Non-isomorphism can be addressed by determining the phases using the multiwavelength anomalous dispersion (MAD) method. These phases can be greatly improved by combining phases from MAD experiments done on different heavy-atom derivatives. Heavy-atom derivatives can be created by chemically modifying the RNA through covalent attachment of bromine or mercury to C5 of pyrimidines or [Pt(NH3)3]2+ to N7 of guanine. While phosphorothioates can provide mercury binding sites, disorder can reduce their value for phase determination. The location of these chemical modifications is critical since crystallization of these derivatized RNAs is sensitive to heavy atom induced conformational alterations and crystal packing.
Correll
C C
CC
Howard Hughes Medical Institute, Yale University, New Haven, CT 06520-8114, USA.
Freeborn
B
B
Moore
P B
PB
Steitz
T A
TA
D001483:Base Sequence; D001966:Bromine; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D008628:Mercury; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010984:Platinum; D012341:RNA, Ribosomal, 5S / Q000737:chemistry*; D013873:Thionucleotides / Q000737:chemistry*
D012341:RNA, Ribosomal, 5S; D013873:Thionucleotides; D010984:Platinum; D008628:Mercury; D001966:Bromine