Translation inhibitors stabilize Escherichia coli mRNAs independently of ribosome protection.
Translation inhibitors such as chloramphenicol in prokaryotes or cycloheximide in eukaryotes stabilize many or most cellular mRNAs. In Escherichia coli, this stabilization is ascribed generally to the shielding of mRNAs by stalled ribosomes. To evaluate this interpretation, we examine here how inhibitors affect the stabilities of two untranslated RNAs, i.e., an engineered lacZ mRNA lacking a ribosome binding site, and a small regulatory RNA, RNAI. Whether they block elongation or initiation, all translation inhibitors tested stabilized these RNAs, indicating that stabilization does not necessarily reflect changes in packing or activity of translating ribosomes. Moreover, both the initial RNase E-dependent cleavage of RNAI and lacZ mRNA and the subsequent attack of RNAI by polynucleotide phosphorylase and poly(A)-polymerase were slowed. Among various possible mechanisms for this stabilization, we discuss in particular a passive model. When translation is blocked, rRNA synthesis is known to increase severalfold and rRNA becomes unstable. Meanwhile, the pools of RNase E and polynucleotide phosphorylase, which, in growing cells, are limited because these RNases autoregulate their own synthesis, cannot expand. The processing/degradation of newly synthesized rRNA would then titrate these RNases, causing bulk mRNA stabilization.
Lopez
P J
PJ
Laboratoire de Génétique Moléculaire (Centre National de la Recherche Scientifique, Unité de Recherche Associée 1302), Ecole Normale Supérieure, 46 rue d'Ulm, 75230 Paris Cedex 05, France.
Marchand
I
I
Yarchuk
O
O
Dreyfus
M
M
D002701:Chloramphenicol / Q000494:pharmacology*; D003513:Cycloheximide / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D011500:Protein Synthesis Inhibitors; C042027:RNA I; D012329:RNA, Bacterial; D012333:RNA, Messenger; D002701:Chloramphenicol; D003513:Cycloheximide; D001616:beta-Galactosidase