Localization of the site of cleavage of ribosomal RNA by colicin E3. Placement on the small ribosomal subunit by electron microscopy of antibody--complementary oligodeoxynucleotide complexes.
Colicin E3 is a ribonuclease that inactivates Escherichia coli ribosomes by cleaving the RNA of the small ribosomal subunit after nucleotide 1493. A series of oligodeoxynucleotides that complement 16 S RNA in the region of the colicin cleavage site has been synthesized, and their ability to form complexes with 30 S ribosomal subunits has been measured using a nitrocellulose filter-binding assay. The most efficiently bound probe, complementary to residues 1485-1496, was modified with antibody-recognizable derivatives at the 5'-end, the 3'-end, or both. Antibody-oligonucleotide-subunit complexes were then generated and examined by electron microscopy. Antibody binding was seen at the tip of the platform of the 30 S subunit. The complementary oligonucleotide and thus the site at which colcin E3 cleavage occurs is therefore in the same physical region as the 3'-end of the 16 S ribosomal RNA and its message-positioning "Shine-Dal-garno" sequence.
Lasater
L S
LS
Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024-1737.
Cann
P A
PA
Glitz
D G
DG
D000906:Antibodies; D000936:Antigen-Antibody Complex / Q000032:analysis; D001483:Base Sequence; D003087:Colicins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D012336:RNA, Ribosomal, 16S / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism / Q000648:ultrastructure; D013379:Substrate Specificity
D000906:Antibodies; D000936:Antigen-Antibody Complex; D003087:Colicins; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D012260:Ribonucleases