G. Garza-ramos et al., Binding site of macrolide antibiotics on the ribosome: New resistance mutation identifies a specific interaction of ketolides with rRNA, J BACT, 183(23), 2001, pp. 6898-6907
Macrolides represent a clinically important class of antibiotics that block
protein synthesis by interacting with the large ribosomal subunit. The mac
rolide binding site is composed primarily of rRNA. However, the mode of int
eraction of macrolides with rRNA and the exact location of the drug binding
site have yet to be described. A new class of macrolide antibiotics, known
as ketolides, show improved activity against organisms that have developed
resistance to previously used macrolides. The biochemical reasons for incr
eased potency of ketolides remain unknown. Here we describe the first mutat
ion that confers resistance to ketolide antibiotics while leaving cells sen
sitive to other types of macrolides. A transition of U to C at position 260
9 of 23S rRNA rendered E. coli cells resistant to two different types of ke
tolides, telithromycin and ABT-773, but increased slightly the sensitivity
to erythromycin, azithromycin, and a cladinose-containing derivative of tel
ithromycin. Ribosomes isolated from the mutant cells had reduced affinity f
or ketolides, while their affinity for erythromycin was not diminished. Pos
sible direct interaction of ketolides with position 2609 in 23S rRNA was fu
rther confirmed by RNA footprinting. The newly isolated ketolide-resistance
mutation, as well as 23S rRNA positions shown previously to be involved in
interaction with macrolide antibiotics, have been modeled in the crystallo
graphic structure of the large ribosomal subunit. The location of the macro
lide binding site in the nascent peptide exit tunnel at some distance from
the peptidyl transferase center agrees with the proposed model of macrolide
inhibitory action and explains the dominant nature of macrolide resistance
mutations. Spatial separation of the rRNA residues involved in universal c
ontacts with macrolides from those believed to participate in structure-spe
cific interactions with ketolides provides the structural basis for the imp
roved activity of the broader spectrum group of macrolide antibiotics.