Bacterial Gene Expression

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Gene Expression Overview - Genes to Proteins

  • Central Dogma (Bacteria): DNA $\xrightarrow{\text{Transcription}}$ RNA $\xrightarrow{\text{Translation}}$ Protein.
    • Pathway: genetic info to functional proteins.
  • Key Molecules & Structures:
    • DNA: Double-stranded; stores genetic code.
    • RNA: Single-stranded; key types:
      • mRNA (messenger): Carries genetic code from DNA to ribosomes.
      • tRNA (transfer): Delivers specific amino acids to ribosomes.
      • rRNA (ribosomal): Major component of ribosomes; catalytic activity.
    • Proteins: Functional molecules (e.g., enzymes, structural elements).
    • Ribosomes: Cellular machinery for protein synthesis (translation).
    • RNA Polymerase: Enzyme catalyzing transcription (DNA $\rightarrow$ RNA).
  • Prokaryotic Hallmark: Transcription and translation are coupled (occur simultaneously in cytoplasm). Coupled transcription and translation in bacteria

⭐ Bacterial mRNA: often polycistronic (one mRNA codes multiple proteins), short half-life; facilitates rapid environmental response.

Bacterial Transcription - Making the Message

Bacterial Transcription: DNA template → mRNA synthesis (5'→3').

  • Enzyme: DNA-dependent RNA Polymerase (RNAP).
      • Holoenzyme: Core enzyme (α₂ββ'ω) + Sigma (σ) factor.
      • Sigma (σ) factor: Recognizes promoter, initiates transcription. Different σ factors for different gene sets (e.g., σ⁷⁰ for housekeeping).
  • Promoter Elements (E. coli):
      • Pribnow box (-10): TATAAT
      • -35 sequence: TTGACA
      • 📌 Mnemonic: "TTGACA" for -35, "TATAAT" for -10.
  • Termination:
      • Rho-dependent: Requires Rho protein (ATPase with helicase activity).
      • Rho-independent (Intrinsic): GC-rich hairpin loop followed by a U-rich sequence in RNA transcript.

Bacterial RNA Polymerase & Promoter Binding

⭐ Rifampicin inhibits bacterial transcription by binding to the β-subunit of RNA polymerase, preventing initiation.

Bacterial Translation - Building the Blocks

  • Ribosomes: 70S (30S + 50S). Site of protein synthesis.
    • 30S subunit: Binds mRNA (via Shine-Dalgarno sequence upstream of AUG). Decodes mRNA.
    • 50S subunit: Contains peptidyltransferase center (23S rRNA - a ribozyme); catalyzes peptide bond formation.
  • Genetic Code: Triplet codons, non-overlapping, degenerate. Start: AUG (codes for N-formylmethionine - fMet in bacteria). Stop: UAA, UAG, UGA.
  • tRNA: Adaptor molecule. Anticodon pairs with mRNA codon. Aminoacyl-tRNA synthetase ensures correct amino acid loading (charging).
  • Process:
    • 📌 Mnemonic for tRNA sites: A (Aminoacyl - Arrives) → P (Peptidyl - Peptide bond) → E (Exit). Bacterial Ribosome Translation Cycle & Antibiotic Targets

⭐ Many antibiotics target bacterial translation: e.g., Aminoglycosides (Streptomycin) bind 30S causing misreading; Macrolides (Erythromycin) bind 50S blocking translocation.

Gene Regulation - Smart Switching

  • Bacteria use operons to regulate gene expression for adaptation.

  • Operon: Promoter, Operator, Structural genes. Regulator gene → repressor.

  • Lac Operon (Inducible - default OFF):

    • Lactose metabolism. Inducer: Allolactose.
    • No lactose: Repressor active, binds operator → transcription OFF.
    • Lactose: Allolactose inactivates repressor → transcription ON.
    • 📌 LACtose Is Needed (for INduction).
  • Trp Operon (Repressible - default ON):

    • Tryptophan synthesis. Corepressor: Tryptophan.
    • Trp low: Repressor inactive → transcription ON.
    • Trp high: Trp activates repressor, binds operator → transcription OFF.
  • Catabolite Repression (Lac operon):

    • Glucose preferred.
    • ↑Glucose → ↓cAMP → CAP inactive → ↓Lac transcription.
    • ↓Glucose → ↑cAMP → CAP active → ↑Lac transcription.

⭐ Glucose inhibits adenylate cyclase, ↓cAMP. cAMP is vital for CAP binding to enhance Lac operon transcription.

Lac Operon Regulation

High‑Yield Points - ⚡ Biggest Takeaways

  • Operons (e.g., lac, trp) are key for coordinated gene regulation in bacteria.
  • Sigma factors direct RNA polymerase to specific promoters, initiating transcription.
  • Coupled transcription-translation in the cytoplasm enables rapid cellular responses.
  • Polycistronic mRNA encodes multiple proteins from one transcript, ensuring efficient pathway synthesis.
  • Regulation primarily occurs at transcription initiation via inducers (e.g., allolactose) and repressors.
  • Catabolite repression (glucose effect) prioritizes efficient energy sources, a global control mechanism.

Practice Questions: Bacterial Gene Expression

Test your understanding with these related questions

Which is the correct sequence of steps in isolating desirable protein using recombinant DNA technology? 1. Expression of protein and lysis of the bacterial cell 2. Incorporation of genes into bacteria 3. SDS PAGE 4. Protein elution 5. Column chromatography

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Flashcards: Bacterial Gene Expression

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Which 5 bacterial toxins depend on lysogeny in order to be secreted by their corresponding bacteria?_____Botulinum ToxinCholera ToxinDiphtheria ToxinShiga Toxin

TAP TO REVEAL ANSWER

Which 5 bacterial toxins depend on lysogeny in order to be secreted by their corresponding bacteria?_____Botulinum ToxinCholera ToxinDiphtheria ToxinShiga Toxin

Group A strep Erythrogenic toxin (causes scarlet fever)

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