Regulation of ribonuclease E activity by the L4 ribosomal protein of Escherichia coli.
Whereas ribosomal proteins (r-proteins) are known primarily as components of the translational machinery, certain of these r-proteins have been found to also have extraribosomal functions. Here we report the novel ability of an r-protein, L4, to regulate RNA degradation in Escherichia coli. We show by affinity purification, immunoprecipitation analysis, and E. coli two-hybrid screening that L4 interacts with a site outside of the catalytic domain of RNase E to regulate the endoribonucleolytic functions of the enzyme, thus inhibiting RNase E-specific cleavage in vitro, stabilizing mRNAs targeted by RNase E in vivo, and controlling plasmid DNA replication by stabilizing an antisense regulatory RNA normally attacked by RNase E. Broader effects of the L4-RNase E interaction on E. coli transcripts were shown by DNA microarray analysis, which revealed changes in the abundance of 65 mRNAs encoding the stress response proteins HslO, Lon, CstA, YjiY, and YaeL, as well as proteins involved in carbohydrate and amino acid metabolism and transport, transcription/translation, and DNA/RNA synthesis. Analysis of mRNA stability showed that the half lives of stress-responsive transcripts were increased by ectopic expression of L4, which normally increases along with other r-proteins in E. coli under stress conditions, and also by inactivation of RNase E. Our finding that L4 can inhibit RNase E-dependent decay may account at least in part for the elevated production of stress-induced proteins during bacterial adaptation to adverse environments.
Singh
Dharam
D
Institute of Molecular Biology, Academia Sinica, Taipei 11529, Taiwan.
Chang
Ssu-Jean
SJ
Lin
Pei-Hsun
PH
Averina
Olga V
OV
Kaberdin
Vladimir R
VR
Lin-Chao
Sue
S
D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D020871:RNA Stability; D012329:RNA, Bacterial / Q000378:metabolism; D012269:Ribosomal Proteins / Q000502:physiology*
D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D012269:Ribosomal Proteins; C056368:ribosomal protein L4; D004722:Endoribonucleases; C022537:ribonuclease E