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Bases
- Purines = double ring structure
o Adenine
o Guanine
- Pyrimidines = single ring structure
o Thymine
o Cytosine
DNA Structure
- Numbering of carbon atoms
- Each strand composed of backbone of alternating
deoxyribose and phosphate molecules
o Held together by covalent bonds (phosphodiester
linkage)
o Between OH group of 3’ carbon of deoxyribose
and phosphate group attached to 5’ carbon
o Leaves 5’ carbon and 3’ carbon free at each end
- Backbones antiparallel (5’ – 3’ vs 3’ - 5’)
- Purines pair with pyrimidines
o AT = 2 H bonds
o CG = 3 H bonds
DNA Packaging
- Nucleosomes
o Consists of two molecules each of four histone proteins (so 8 histone proteins),
forming the nucleosome core
o DNA wraps twice around these eight protein molecules
o Fifth type of histone protein helps maintain the nucleosome
DNA Sequences
- Only 2% of DNA is coding DNA (codes for proteins)
- Rest of DNA:
o Acts as regulators of gene expression
o Introns (junk DNA)
o Telomeres
o Codes for tRNA
- Highly repetitive sequences
o Usually comprised of 5-300 base pairs
o Clustered repetitive DNA = satellite DNA
o Transposons = repetitive sequences that can change position within the DNA
(regulators of gene expression)
- Protein coding genes
o Genes that have coding functions
o Base sequence carried from nucleus to ribosome by mRNA
o Composed of exons (coding) and introns (non-coding)
- Structural DNA
o Highly coiled DNA
o Does not have a coding function
o Occurs around centromere and telomeres (ends of each chromosome)
- Short tandem repeats (STRs)
o Short (2-5 bases) repeating sequences of DNA
o Most DNA is identical, repeating sequences that show variation between people
= polymorphisms
o Polymorphisms used for DNA profiling
o Scientists use 13 different loci for DNA profiling called STRs
DNA Replication
- Begins at sites called origins of replication
o Prokaryotic DNA has single origin
o Eukaryotic DNA has thousands of origins
- Summary:
o Begins at origin which appears as a bubble (unzipping catalyzed by helicase
breaking H bonds)
o At the end of each bubble, there is a replication fork (where double-stranded
DNA opens to provide two parental DNA template molecules)
o Bubbles enlarge in both directions; replication process is bidirectional
o Bubbles fuse to produce two identical daughter DNA strands
- Elongation of a new DNA strand
o Primer produced under direction of primase at replication fork
Primer = short sequence of RNA (5 – 10 bases long)
Primase allows joining of RNA nucleotides that match exposed DNA bases
at point of replication
o DNA polymerase III allows addition of DNA nucleotides in 5’ to 3’ direction to
produce growing DNA strand
Nucleotide = dNTP (deoxynucleoside triphosphate)
Contains deoxyribose, nitrogenous base, and 3 phosphate molecules
Loss of 2 phosphate molecules energy provided for bonding to occur
o DNA polymerase I removes primer from 5’ end and replaces it with DNA
molecules
- Antiparallel strands
o Strands run in 5’ to 3’ and 3’ to 5’ direction antiparallel
o DNA polymerase III assembles in 5’ to 3’ direction on daughter strand
o Leading strand and lagging strand assembled concurrently
o Lagging strand forms more slowly, involved Okazaki fragments, and DNA ligase
Leading strand formed from 5’ to 3’ continuously towards progressing
replication fork
Lagging strand assembled by Okazaki forks moving away from
progressing replication fork (5’ to 3’)
Primer, primase, and DNA polymerase III required to begin formation of
each Okazaki fragment and to begin formation of leading strand
Primer and primase only needed once for leading strand
DNA ligase attaches sugar-phosphate backbones of fragments to form a
single DNA strand
- Replication proteins
Transcription
- RNA polymerase separates two DNA strands,
allows polymerization of RNA nucleotides
o 5’ 3’ direction
- DNA strand that carries genetic code is sense
strand (coding strand) vs. antisense strand
(template strand)
o ANTISENSE strand is copied