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Ch. 12 B
Gene structure, replication, and expression
83
Biology
Undergraduate 3
03/05/2011

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Term
The Genetic Code
Definition
  • Only about 20 AA produced by sequence of 3 bases in mRNA called a codon
Term
Code Degeneracy
Definition
  • Several Codons call for the same AA to be used during translation
    • ie: CUU, CUC, CUA, CUG encode leucine
Term
In a code degeneracy:
Definition
  • There is only 1 start codon: AUG (in most prokaryotes)
Term
3 stop/Nonsens codons
Definition
  • UAA
  • UAG
  • UGA
Term
Codon Translation
Definition
Translacted by complementary base-pairing of tRNA anticodon
Term
In Codon translation 1:
Definition
  • tRNAs aminoacetlyated: each carries specific amino acid
  • Anticodon diversity< codon diversity
Term
What is the WOBBLE hypothesis?
Definition
Terminal base of anticodon is replaced with inosine
Term
What can inosine pair with?
Definition
  • Cytosine, uracil, or adenine (allows for degeneracy)
Term
What is needed in prokaryotice translation?
Definition
All 3 forms of prokaryotic RNA essential
Term
Why is RNA essential to Prokaryotic Translation?
Definition
  • it plus proteins form ribosome structure and translation occurs within this complex
Term
Why is tRNA essential to prokaryotic translation?
Definition
it brings in AA to be added to polypeptide chain
Term
Why is mRNA essential to Prokaryotic Translation?
Definition
  • Serves as recipe for polypeptide chain
Term
Polypeptide Synthesis
Definition
The free C-terminal (-COOH group) of first amino acid is joined to N-terminal (NH2- group) of the next amino acid via peptide bond
Term
Comparison of polypeptide and mRNA synthesis
Definition
  • Polypeptide is SLOWER than mRNA synthesis
    • transcription 900 residues per min
    • translation 110 residues per min
Term
What does polypeptide synthesis do to translation efficiency?
Definition
  • Increases it by using polysomes
    • More than 1 ribosome translates the mRNA at same time
    • Results in multiple copies of polypeptide being produces per mRNA molecule.
Term
Prokaryotic Ribosome
Definition
  • 70S whole ribosome (50S and 30 S subunits)
Term
Subunits of Prok. Ribosomes
Definition
Remain separate in cyto until mRNA is avaliable
Term
Translational domain area of prok. ribosomes:
Definition
Site where polypeptide synthesis actively occurs; formed by both subunits of ribosome
Term
How is mRNA aligned to translational domain?
Definition
through complementary binding of mRNA leader sequences (Shine-Delgarno) to specific regions near translational domain of mRNA
Term
Exit domain of Pro. Ribosomes
Definition
During translation, nascent protein forms and emerges from ribosome at this part
Term
Steps in Translation: Initiation 1
Definition

f- Met carried into ribosome by tRNA and binds to free 30 S subunit of rRNA

  • positioned at what will become P-site of Active Ribosome Complex
Term
What will happen after translation to the formyl group?
Definition
It can be cleaved off to render a plain methionine residue
Term
Steps in Translation: Initiation 2
Definition

mRNA attaches to 30S subunit

  • It allighns itself so AUG intiator godon is @ translation domain of 30S subunit site
  • Initiator codon complexes with complementary tRNA anti-codon (UAC)
Term
What helps mRNA position @ the translator domain of 30 S subunit site during initation of translation?
Definition
Shine Delgarno
Term
Steps in Intiation: Translation 3
Definition

Complete active ribosome complex forms

  • mRNA binds to 50S subunit
  • Energy required: GTP--> GDP + inorganic Phosphate (1ATP equivalent)
Term
IF-1 in bacteria (functions)
Definition
  • Helps bind 30S and 50S subunits together
  • Helps release IF-2/GDP from ribosome
  • Blocks tRNA from binding to A site (aminoacyl site, also known as acceptor site)
Term
IF-2
Definition
  • Used as an energy source with GTP (GDP produced)
  • Makes sure fMet tRNA binds to P site
Term
IF-3
Definition
Prevents 30 S and 50S subunits from binding to one another until mRNA has complexed with 30 S subunit
Term
How much GTP is required to form an active ribosome complex?
Definition
1
Term
Elongation step 1:
Definition
Next appropriate aminoacyl-tRNA binds to ribosome in A-site (aminoacyl/ acceptor)
Term
How does the elongation step occur?
Definition
via complementary condon-anti codon binding
Term
Does step 1 of elongation require energy?
Definition
Yes, GTP--> GDP + Inorganic Phosphate (1 ATP equivalent)
Term
Elongation Step 2
Definition

Transpeptidation occures between adjacent amino acids in P and A sites

 

Term
Step 2 of elongation causes:
Definition
  • Elongation of Polypeptide chain
  • Peptidyl transferase creates peptide bond between adjacent amino acids
Term
Step 3 elongation
Definition
Translocation of ribosome occurs
Term
Where is A site in Elongation?
Definition
Rests over a new open codon
Term
Where does the growing polypeptide chain move to during elongation?
Definition

the P-site

  • Long polypep begin to exit ribosomes translational domain via e-site
  • tRNA which have donated their amino acid also exit ribosome via e-site
Term
Is there energy required in step 3 of elongation?
Definition
Yes, GTP--> GDP + inorganic phosphate (1ATP equiv)
Term
Elongation step 4
Definition
Repeat 1-3 until stop codon positioned at A site
Term
P-site in elongation
Definition
  • Only tRNA carring N-fmet binds here during initiation
  • During elongation, growing chain is moved here
Term
A-Site
Definition
  • Empty at start of translation
  • Fills during elongation
Term
EF-Tu (w/GTP)
Definition
Binds an aminoacyl tRNA into A site
Term
EF-G (w/GTP)
Definition
  • Also called translocase
  • GTP used to 'move' the ribosome to next codon
Term
Summary of Elongation
Definition
  • Require 1 GTP to bind to aminoacyl-tRNA in A site
  • Require 1 GTP to translocate ribosome
  • 2 GTP/codon of elongation
Term
Step 1 Termination
Definition
Ribosome translocated to stop/ nonsense codon
Term
What exactly happens in step 1 of termination?
Definition
  • No tRNA anticodons avaliable to undergo complementary binding to mRNA stop codons
  • Translation stalls out
Term
Step 2 Termination
Definition
Peptidy transferase hydrolyzes last bond between t-RNA (in P site) and growing pep. chain
Term
What exactly does step 2 Termination do?
Definition
  • Frees polypeptide and tRNA to leave ribosome
Term
Is energy used in step 2 of termination?
Definition
Yes GTP--> GDP +Pi (1 ATP equivalent)
Term
Step 3 Termination
Definition

Ribosome subunits dissociate from 1 another and from mRNA

  • Subunits remain as free entities in cyto
  • IF-3 rebinds to 30 S subunit to keep them apart again.
Term
Termination step 4
Definition

Other ribosomes of polysome finish translating mRNA to stop codon

  • After last ribosome finishes, mRNA can be degraded back to individual nucleotides by cytoplasmic nucleases
Term
RF-1, RF-2, and RF-3
Definition

aid ribosome in recognizing the stop codons

 

specific roles currently undetermined

Term
Release factors
Definition
use 1 GTP molecule to release ribosome units from mRNA and each other
Term
Cost of Protein synthesis:
Definition
  • 1 GTP to bind the initial N-fmet to P site
  • 2 GTP/codon during elongation and translocation
  • 1 GTP at termination
Term
@ Cost of Protein sythesis, what would happen
Definition
All of cells energy would be expended by producing both essential and non-vital proteins
Term
How can energy be saved in protein synthesis?
Definition
Regulating gene expression for proteins not in constant demand
Term
How do protiens mature in bacteria?
Definition

Native proteins under go folding

  1. Exits ribosome in promary structure
  2. folds into secondary and tertiary shapes
  3. Joins other tert. proteins to produce quarternary structures
Term
What assists protein maturation in bacteria?
Definition
Special chaperone proteins
Term
What are some examples of special chaperone proteins?
Definition
  • DNaK and DnaJ
  • GroEL and Gro ES
  • GrpE
Term
  • DNaK and DnaJ
Definition
begin folding for native proteins and use ATP
Term

 

  • GroEL and Gro ES
Definition
Completes folding into functional proteins using more ATP
Term
GrpE
Definition
known as stress protein, involved in heat shock survival
Term
Secretion
Definition
when cells must excrete certain proteins to outside environment
Term
Secretion of Gram +
Definition
Only need to transport through plasma membrane
Term
Secretion of Gram Neg
Definition
Bac. must be transported through both the plasma and the outer
Term
All Sercretion pathways require what?
Definition
Energy
Term
What are the two energy sources in the pathway of secretion?
Definition
  1. ATP/GTP hydrolysis
  2. Proton Motive Force
Term
Sec Dependent Secretion
Definition
  • Occurs in Gram + and Gram -
  • Transports only across cytoplasmic mem
  • Transported as pre-proteins w/ single pep at N-term and then sec B chaperone recognizes signal
Term
Type I (ABC transport)
Definition
  • Gram - and Gram + Bacteria
    • C term used for recog of new proteins
    • Spans cyto and outer membrane
  • No sec-dependent transport needed
  • Transports things other than proteins
  • expels drugs from cyto
Term
Type II
Definition
  • Gram - only
  • Common in pathogens
  • Transports : cellulases, pectinases, proteases, lipases, toxins across outher membrane from periplasmic space by Sec-dependent and Tat pathways
Term
How are Tat and sec different?
Definition
Tat secretes folded proteins
Term
Type V
Definition
  • Gram - only
  • Transports proteins periplasmic--> outer membrane
  • uses sec-dependent
  • some autotransporters, some require helper protein
Term
Type III
Definition
  • Gram - bacteria only
  • Transports directly cyto--> cell exterior
  • No sec needed
  • Transports: toxins, phagocytosis inhibitors, apotopsis promoters, secretion reg. proteins
Term
Type IV
Definition
  • Gram + and Gram - Bacteria
  • Secretes both proteins and DNA from cyto--> outside cell
Term
Antibiotics and protein synthesis:
Definition
  • Binding to 30S or 50S subunit- prevents active ribosome mRNA complex from ever forming and hence, prevent translation
  • Interfering w/ ribosomes ability to bind to DNA, thus prevents transcription
Term
How does Antibiotics prevent translation?
Definition
  • Binding to 30S or 50S subunit- prevents active ribosome mRNA complex from ever forming
Term
How do Antibiotics prevent transcription?
Definition
  • Interfering w/ ribosomes ability to bind to DNA
Term
Selective toxicity for bacteria
Definition

Eukaryotes have different types of rRNA subunits and RNA polymerases

  • prokaryotic seq. in rRNA recognizable by antis not present in euk 
  • Different polymerases
Term
What does the selective toxicity for bacteria cause?
Definition
Inhibits anitibiotics harmless to us, but deadly to the bacteria affecting us.
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