The A2453-C2499 wobble base pair in Escherichia coli 23S ribosomal RNA is responsible for pH sensitivity of the peptidyltransferase active site conformation.
Peptide bond formation, catalyzed by the ribosomal peptidyltransferase, has long been known to be sensitive to monovalent cation concentrations and pH. More recently, we and others have shown that residue A2451 in the peptidyltransferase center of the Escherichia coli 50S ribosomal subunit changes conformation in response to alterations in pH, depending on ionic conditions and temperature. Two wobble pairs, A2453-C2499 and A2450-C2063, have been proposed as potential candidates to convey pH-dependent flexibility to the peptidyltransferase center. Each is presumed to possess a near-neutral pK(a), and both lie in proximity to A2451. We show through mutagenesis and chemical probing that the identity of the A2453-C2499 base pair, but not the A2450-C2063 base pair, is critical for the pH-dependent structural rearrangement of A2451. We conclude that, while the A2453-C2499 base pair may be important for maintaining the structure of the active site in the E.coli peptidyltransferase center, its lack of conservation makes it, and consequently its near-neutral pK(a), unlikely to contribute to function during peptide bond formation.
Bayfield
Mark A
MA
Department of Molecular Biology, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
Thompson
Jill
J
Dahlberg
Albert E
AE
D000225:Adenine / Q000737:chemistry; D020029:Base Pairing; D001665:Binding Sites; D003596:Cytosine / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D010458:Peptidyl Transferases / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000235:genetics; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D013463:Sulfuric Acid Esters / Q000737:chemistry
D012338:RNA, Ribosomal, 23S; D013463:Sulfuric Acid Esters; D003596:Cytosine; D010458:Peptidyl Transferases; D000225:Adenine; C007482:dimethyl sulfate