Recombinant Escherichia coli GTPase Era (era)
Description
Functional Roles in Ribosome Biogenesis
Era orchestrates critical steps in 30S ribosomal subunit maturation:
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16S rRNA Processing: Binds precursor 16S rRNA near its 3′ end, facilitating endonucleolytic cleavage by YbeY and exonucleolytic trimming by RNase II, R, PH, and PNPase .
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Quality Control: Ensures proper folding of the 30S platform region, preventing misfolded intermediates .
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GTPase Activity: Hydrolyzes GTP to GDP, with activity stimulated by RNA binding .
Table 2: Functional Characteristics
3.1. Mechanism of GTPase Activation
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RNA binding (e.g., 16S rRNA) enhances GTP hydrolysis by stabilizing Era’s active conformation .
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Critical residues:
3.2. Suppression of Ribosome Defects
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Overexpression of Era rescues growth defects in ΔybeY strains by improving 16S rRNA processing and 70S assembly .
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GroEL interaction: Era(T99I) increases GroEL association with 30S subunits, suggesting chaperone-mediated stabilization .
3.3. Evolutionary Conservation
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Era homologs exist in mitochondria (ERAL1) and chloroplasts, where they maintain roles in ribosome assembly .
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Human disease link: ERAL1 mutations cause Perrault syndrome, highlighting Era’s conserved importance .
Recombinant Era in Experimental Systems
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Expression systems: E. coli BL21(DE3) commonly used for His-tagged or GST-fusion Era .
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Purification: Affinity chromatography (Ni-NTA or glutathione resins) followed by RNA removal via ion exchange .
Table 3: Mutational Analysis of Era
Regulatory and Interaction Networks
Product Specs
Target Background
KEGG: ecm:EcSMS35_2719
