Recombinant Elusimicrobium minutum Translation initiation factor IF-3 (infC)
Description
Introduction to Translation Initiation Factor IF-3
Translation initiation factor IF-3 is a critical component of prokaryotic translation machinery, ensuring fidelity during ribosome assembly and start codon selection. In bacteria, IF3 prevents premature 50S subunit binding to the 30S pre-initiation complex (PIC), verifies codon-anticodon pairing, and discriminates against non-initiator tRNAs . Structurally, IF3 comprises two domains:
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N-terminal domain (NTD): Modulates interactions with initiator tRNA (i-tRNA) and cooperates with the C-terminal domain (CTD) for ribosomal subunit dissociation.
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C-terminal domain (CTD): Directly prevents 50S subunit binding and stabilizes i-tRNA in the P-site .
Elusimicrobium minutum IF-3 (infC) Genomic Context
Elusimicrobium minutum, a member of the candidate phylum Elusimicrobia, is a strict anaerobe first isolated from beetle larvae guts. Its genome (1.64 Mbp, 39% GC content) encodes 1,529 protein-coding genes, including essential translation machinery components . While direct annotation of infC (the gene encoding IF-3) is not explicitly detailed in available genomic studies, comparative analyses suggest its presence due to:
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Conservation of translation initiation factors across bacteria.
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Genomic evidence for ribosomal proteins and tRNA modification enzymes .
A hypothetical domain structure of E. minutum IF-3, inferred from homologs like Escherichia coli IF-3, is provided below:
Recombinant Expression of E. minutum IF-3
Recombinant IF-3 production typically involves cloning the infC gene into expression vectors (e.g., pET or pQE systems) under inducible promoters (e.g., T7 or araBAD) . Key steps include:
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Gene Synthesis: Codon optimization for expression in E. coli or other hosts.
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Vector Design: Inclusion of affinity tags (e.g., His-tag) for purification .
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Functional Validation:
Future Directions
Product Specs
Target Background
KEGG: emi:Emin_0346
STRING: 445932.Emin_0346
Q&A
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What experimental designs are effective for studying the function of the Translation initiation factor IF-3 in Elusimicrobium minutum?
To study the function of the Translation initiation factor IF-3 in Elusimicrobium minutum, researchers can employ a combination of genetic manipulation and biochemical assays. A common approach involves creating knockout strains using CRISPR-Cas9 technology to assess the impact of IF-3 deletion on protein synthesis and growth rates. Additionally, in vitro translation assays can be conducted to evaluate the role of IF-3 in ribosome assembly and initiation complex formation. Data from these experiments should be analyzed statistically to determine significance, particularly focusing on variations in growth rates and translation efficiency between wild-type and mutant strains.
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How can data contradictions in studies of Elusimicrobium minutum be addressed through systematic reviews?
Systematic reviews can help reconcile contradictions in research findings related to Elusimicrobium minutum by synthesizing data across multiple studies. Researchers should establish clear inclusion criteria for relevant studies, focusing on those that investigate aspects such as metabolic pathways or genetic characteristics. A meta-analysis can be performed to quantitatively assess discrepancies, allowing for the identification of potential biases or methodological flaws in individual studies. This comprehensive approach not only clarifies existing contradictions but also highlights areas requiring further investigation.
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What advanced techniques are available for analyzing the metabolic pathways of Elusimicrobium minutum, particularly concerning its Translation initiation factor IF-3?
Advanced techniques such as metabolomics and transcriptomics are crucial for analyzing the metabolic pathways associated with Elusimicrobium minutum and its Translation initiation factor IF-3. Metabolomic analysis involves using mass spectrometry or nuclear magnetic resonance spectroscopy to profile metabolites produced under varying conditions, providing insights into metabolic fluxes influenced by IF-3 activity. Transcriptomic studies, utilizing RNA sequencing, can reveal differential gene expression patterns linked to translation initiation processes. Integrating these approaches allows for a holistic understanding of how IF-3 impacts metabolic regulation.
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What role does the genomic structure of Elusimicrobium minutum play in its translation initiation mechanisms?
The genomic structure of Elusimicrobium minutum, characterized by a relatively small circular chromosome with specific gene arrangements, influences its translation initiation mechanisms significantly. The presence of genes encoding ribosomal proteins and translation factors, including IF-3, suggests a streamlined process tailored to its anaerobic lifestyle. Researchers can analyze gene synteny and regulatory elements through comparative genomics with closely related species to elucidate how genomic features facilitate efficient protein synthesis under anaerobic conditions.
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How does the phylogenetic position of Elusimicrobium minutum inform our understanding of its translation initiation factors?
The phylogenetic position of Elusimicrobium minutum within the TG1 phylum provides insights into the evolutionary adaptations of its translation initiation factors, including IF-3. By comparing its genomic sequences with those from other phyla, researchers can identify conserved domains and unique adaptations that may enhance translation efficiency in extreme environments, such as those found in insect guts. Phylogenetic analyses can reveal evolutionary pressures that shaped the functional characteristics of these factors, guiding future research on their roles across different microbial communities.
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What methodologies are recommended for assessing the functional interactions between Translation initiation factor IF-3 and ribosomal RNA in Elusimicrobium minutum?
To assess functional interactions between Translation initiation factor IF-3 and ribosomal RNA in Elusimicrobium minutum, researchers should consider employing co-immunoprecipitation followed by mass spectrometry to identify binding partners. Additionally, fluorescence resonance energy transfer (FRET) assays can provide real-time insights into interactions during translation initiation. These methodologies allow for a detailed understanding of how IF-3 interacts with ribosomal RNA and other components during protein synthesis, contributing to our knowledge of translational regulation in this organism.
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What are the implications of studying Translation initiation factor IF-3 for understanding microbial ecology in gut environments?
Investigating Translation initiation factor IF-3's role in Elusimicrobium minutum has broader implications for microbial ecology within gut environments. Understanding how this factor influences protein synthesis can shed light on competitive dynamics among gut microbiota, particularly regarding nutrient utilization and adaptation strategies under anaerobic conditions. Researchers could use metagenomic approaches to correlate the presence of specific translation factors with community composition and functionality, providing insights into microbial interactions and their ecological significance.
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How does the presence of hypothetical genes in the genome of Elusimicrobium minutum impact research on Translation initiation factors?
The presence of numerous hypothetical genes in the genome of Elusimicrobium minutum presents both challenges and opportunities for research on Translation initiation factors like IF-3. These genes may encode novel proteins that interact with known translational machinery or participate in regulatory networks affecting translation efficiency. Functional characterization through gene knockout or overexpression studies is essential to determine their roles, potentially leading to discoveries that enhance our understanding of translational control mechanisms in this organism and similar bacteria.
