PMID:2105934

Citation	Chen, SM, Takiff, HE, Barber, AM, Dubois, GC, Bardwell, JC and Court, DL (1990) Expression and characterization of RNase III and Era proteins. Products of the rnc operon of Escherichia coli. J. Biol. Chem. 265:2888-95
Abstract	The synthesis rates of ribonuclease III (RNase III) and Era proteins are relatively low, and expression of the era gene is translationally coupled with expression of the rnc gene. Expression of both genes is negatively controlled by RNase III itself. We have constructed plasmids that overproduce RNase III and/or Era proteins under the control of the lambda PL promoter. A plasmid with the rnc gene under PL control expresses RNase III at levels greater than 40% of total cellular protein. Another plasmid with the era gene under PL control and a modified translation-initiation signal produces up to 80% of total cell protein as Era. Each protein has been purified using simple and rapid procedures. Purified RNase III protein specifically processes mRNA transcripts containing known RNase III sites. The purified Era protein binds GDP and GTP and has GTPase activity. Kinetic analysis shows that one molecule of GTP or GDP is bound/Era peptide with a Kd of 5.5 microM for GTP binding and 1.0 microM for GDP binding. The Km of the Era GTPase is 9.0 microM, and the maximum catalyzed rate of GTP hydrolyzed/min/mol of Era protein at 37 degrees C is 9.8 mmol.
Links	PubMed
Keywords	Bacterial Proteins/biosynthesis; Bacterial Proteins/genetics; Bacterial Proteins/isolation & purification; Base Sequence; Endoribonucleases/biosynthesis; Endoribonucleases/genetics; Endoribonucleases/isolation & purification; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli Proteins; GTP-Binding Proteins/genetics; GTP-Binding Proteins/metabolism; Gene Expression; Genes, Bacterial; Kinetics; Molecular Sequence Data; Operon; Plasmids; Restriction Mapping; Ribonuclease III; Sequence Homology, Nucleic Acid
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ECOLI:ERA		GO:0003924: GTPase activity	ECO:0000314: direct assay evidence used in manual assertion		F		Figure 8A. shows that soluble forms of Era, when coupled with GTP, yield both GTP and GDP. This is evidence of GTPase activity.	complete CACAO 3826
ECOLI:ERA	enables	GO:0003924: GTPase activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

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