Three-dimensional placement of the conserved 530 loop of 16 S rRNA and of its neighboring components in the 30 S subunit.
Nucleotides 518-533 form a loop in ribosomal 30 S subunits that is almost universally conserved. Both biochemical and genetic evidence clearly implicate the 530 loop in ribosomal function, with respect both to the accuracy control mechanism and to tRNA binding. Here, building on earlier work, we identify proteins and nucleotides (or limited sequences) site-specifically photolabeled by radioactive photolabile oligoDNA probes targeted toward the 530 loop of 30 S subunits. The probes we employ are complementary to 16 S rRNA nucleotides 517-527, and have aryl azides attached to nucleotides complementary to nucleotides 518, 522, and 525-527, positioning the photogenerated nitrene a maximum of 19-26 A from the complemented rRNA base. The crosslinks obtained are used as constraints to revise an earlier model of 30 S structure, using the YAMMP molecular modeling package, and to place the 530 loop region within that structure.
Wang
R
R
Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA.
Alexander
R W
RW
VanLoock
M
M
Vladimirov
S
S
Bukhtiyarov
Y
Y
Harvey
S C
SC
Cooperman
B S
BS
D003432:Cross-Linking Reagents / Q000528:radiation effects; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000648:ultrastructure*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D019920:Photoaffinity Labels / Q000528:radiation effects; D012329:RNA, Bacterial / Q000737:chemistry* / Q000528:radiation effects; D012333:RNA, Messenger / Q000737:chemistry / Q000528:radiation effects; D012336:RNA, Ribosomal, 16S / Q000737:chemistry* / Q000528:radiation effects; D012269:Ribosomal Proteins / Q000737:chemistry / Q000528:radiation effects; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*