Exam 4 Flashcards
DNA was first (1869) isolated by Swiss chemist
Friedrich Meischer; called it “nuclein”.
(1929) Phoebus Levene discovered
deoxyribose as sugar component of DNA.
Discovered four nitrogenous bases of DNA: thymine, adenine, guanine, and cytosine.
Tetranucleotide hypothesis: DNA consisted of short chains of 4 repeating nucleotide units.
(1928-1943) Several investigators reported experimental results suggesting that DNA carried
heritable information
Nucleic acids are polymers of
nucleotides
nucleotides are consisted of
pentose sugar
- ribose in RNA
- deoxyribose in DNA
nitrogenous base consists of
purines and pyrimidines.
Purines: G, A
Pyrimidines: C, T (DNA), U (RNA)
Phosphate group consists of
Nucleoside: nitrogenous base + sugar.
Nucleotide: nucleoside + ≥ 1 phosphate.
Chargaff and Base Composition of DNA
analyzed base composition of DNA from several different species
base comparison varies between species
- suggests molecular diversity
regularity in the ratio of nucleotide bases
Chargaff’s Rules
Rule 1: Base composition of DNA varies btw species.
Rule 2: %A ≈ %T; %G ≈ %C.
- Use to calculate unknown %s.
Exception: фX174 = bacteriophage
- Has single stranded, circular genome.
Rosalind Franklin (7/25/1920 – 4/16/1958)
British biophysicist working in X-ray crystallography.
- ↑ understanding molecular structure of DNA, RNA, viruses.
Data was critical to Watson-Crick DNA model.
- Shown to Watson w/o her knowledge.
- Prepared papers recognizing two forms A and B-DNA
Franklin’s X-ray Crystallography Data
X-rays are deflected as pass through DNA fibers.
Watson and Crick assembled models conforming to Franklin’s data and Chargaff’s rules.
Watson-Crick Model of Double Helix
Duplex DNA: structure consists of two strands.
Two strands are anti-parallel:
Followed Franklin’s conclusion: sugar-phosphate backbone was outside.
- Places hydrophobic bases interior.
Helix makes a full turn every 3.4 nm and bases are stacked 0.34 nm apart.
Nucleic Acids are Directional
Phosphodiester bonds result in a polymer w/ distinct ends.
5’ end: ends in terminal phosphate.
3’ end: ends in pentose sugar with free 3’ hydroxyl group.
Covalently linked sugars and phosphates make up phosphodiester backbone.
________ - ________ only pairing that fits.
Purine-pyrimidine
Strands stabilized by hydrogen bonds.
Explains Chargaff’s Rules
Dictates combos btw strands
No sequence restrictions along strand
Watson-Crick base pairing is key to information sharing in
DNA replication
Transcription (DNA > RNA)
Translation (mRNA > polypeptide [protein])
Leading strand
synthesized continuously in direction of replication fork.
Lagging strand
synthesized discontinuously in direction away from replication fork
Helicase
uses ATP hydrolysis to unwind DNA duplex
Primase
Synthesizes a short RNA primer.
DNA polymerase
DNA-dependent synthesis of comple-mentary DNA molecule.
DNA ligase
joins adjacent replicated strands.
Enzymes that catalyze synthesis of new DNA by adding _________ to a preexisting chain.
nucleotides
Require primer and a template strand.
E. coli
several DNA polymerases; two most important for DNA replication
DNA Pol I: removes RNA primer and replaces w/ DNA.
- DNA Pol III: adds DNA nucleotide to RNA primer and continues to add Nts complementary to template strand.
Rate of elongation
≈ 500 Nt/sec in bacteria and ≈ 50 Nt/sec in human cells.
DNA polymerase
catalyzes addition of a nucleotide to the 3’ end of a growing chain
Each nucleotide added is in the form of dNTP
dNTP
- each monomer is added, pyrophosphate is lost.
- Hydrolysis of pyrophosphate is a coupled exergonic rxn.
Structural differences RNA vs. DNA
Sugar = ribose (2’ OH)
Uracil (U) instead of thymine (T).
Usually single stranded
Messenger RNA
mRNA: carries information from DNA to ribosome
Transfer RNA
tRNA: delivers specific amino acids to ribosome
Ribosomal RNA
rRNA: structural and catalytic part of ribosome.
Regulatory RNA
regulate gene expression; include microRNA (miRNA), long, non-coding RNA (lncRNA), small interfering RNA (siRNA).
intramolecular base pairings of purines and pyrimidines
A=U
G=C
creates intramolecular regions with different structures;
stem: complementary base pairing by antiparallel turns of RNA strand.
loop: unpaired region
(Francois Jacob and Jacques Monod, 1961) hypothesized that single-stranded RNA
messenger RNA carries info from DNA to ribosomes.
Gene Expression in Bacteria
Most genes are continuous.
Some genes are organized into operons.
- Produce a polycistronic mRNA.
Lack of compartment-alization allows coupling of transcription and translation
Template strand
used to direct transcription; complementary and antiparallel to RNA.
- Also called antisense strand.
Non-template strand
NOT usually transcribed; same sequence and polarity as RNA
- contains U instead of T.
- also called sense = coding strand.
Promoter
DNA sequence immediately upstream (5’) of transcription start site (tss);
- Not transcribed; controls binding of RNA polymerase
RNA coding region:
portion of gene transcribed into mRNA
Termination region
sequences located 3’ to coding region; signals end of transcription
A single promoter controls the expression of 3 genes
lacZ, lacY, and lacA
Products of lacZ and lacY genes involved in lactose metabolism
3 clustered genes are coordinately expressed in single mRNA
polycistronic mRNA
uncoupled
Nuclear envelope separates trans-cription and translation
- transcription = nuclear.
- mRNA is transported to cytoplasm for translation.
RNA splicing
introns are removed from pre-mRNA
5’ cap
addition of a modified guanine to 5’ end of mRNA.
- added after ≈ 20-40 Nts are transcribed.
poly-A tail
addition of 50-250 adenine residues of 3’ end of pre-mRNA.
Functions:
- Prevent degradation
- Help mRNA transport
- Help position mRNA on ribosome