Unit 2 Flashcards
Nucleoside
Base connected to sugar
Nucleotide
Base + Sugar + Phosphate
Phosphodiester Bond
Btwn nucleotides
Form backbone of nucleic acids
Covalent
Glycosidic Bond
Connection btwn base and sugar
Covalent
Hydrogen Bond
Form base pairs
Non covalent
Watson Crick Franklin Pairing
Anti/Anti
C+G
A+T
Hoogsteen Pairing
Syn/Anti
Exotic RNA structures
Purines
A and G
2 rings
Pyrimidines
C, T, and U
1 ring
DNA vs RNA
DNA lacks 2’ OH group
Ester Linkage
Attach phosphate to nucleoside
A + T
2 x H-bonds
G + C
3 x H-bonds
Base stacking interactions
van der Waals (btwn base faces)
pi stacking
B form structure
Anti parallel
Right handed
Backbone on outside
Bases on inside
Major groove
Wide and deep
Minor groove
Shallow and narrow
RNA preferred conformer
C3’ endo
C2’ endo causes sterics
DNA preferred conformer
C2’ endo
T melting
Temp at which the helix is 1/2 double stranded and 1/2 single stranded
Negative supercoiling
Lk < 0
DNA = underwound
Positive supercoiling
Lk > 0
DNA = overwound
Topoisomerase
Change DNA topology
Resolve supercoiling that develops when genome is locally unwound
Nucleosome
DNA wrapped around a histone
Chromatin
Histones packaged into 30 nm fiber
Involves supercoiling
Heterochromatin proteins
Bind across methylated histones
Inaccessible
Transcription can’t occur
Marked by methylation
Euchromatin
Accessible
Transcription can occur
Marked by acetylation
Chem mechanism of nucleotide addition
Base activates 3’ OH
3’ OH does nucleophilic attach on 5’ phosphate
Synthesis direction
5’ to 3’
DNA Polymerase
Catalyze nucleotide addition
Requires stretch of primer
Polymerase and Mg2+
Mg2+ facilitates chemistry
- Activates 3’OH
- Stabilize - charge of phosphate
- Mg2+ binds to Asp
Primer
Short stretch of RNA annealed to template
Facilitates addition of complementary base pairs
Takes advantage of energy of base stacking
DNA Polymerase Mistakes (2)
Tautomerization
Wobbles
Wobbles
Normal bases that bind inappropriately
due to shift in position of nucleotides
Tautomerization
Change into form where positions of H-bond donor/acceptor changes
DNA Polymerase Activities
Add nucleotides via polymerase domain
Remove mismatches via exonuclease domain
Incorrect base detection
Destabilization of DNA structure
Adopt conformation that moves 3’ end into exonuclease site
Incorrect base correction
3’ end enters exonuclease site
Terminal nucleotide removed
Polymerase gets 2nd chance
DNA Pol III
Main replicative polymerase
Built for speed
Copies entire genome
Built for processivity
DNA Pol I
Odd jobs
Polymerizes small stretches as part of cleanup/repair
Built for accuracy
Start of replication
Origin of replication
Helicase
Melt and unwind DNA
Primase
Synthesize small RNA sequence as a primer for DNA Pol III
Leading strand
Continuous synthesis
Parent 3’ end on the bottom left
Lagging strand
Discontinuous synthesis
Parent 3’ end on the top right
Okazaki fragments
Fragments that make up discontinuous strand
Lagging strand activities
Primase adds primer
Pol III extends DNA until encounter of next fragment
Primase + Pol III make fragments
Pol I removes primer + replaces with DNA
Ligase seals fragments
Ligase
Seals nicked DNA fragments
How does Ligase work
- Ligase uses ATP to adenylate itself
- AMP transferred to 5’ phosphate of nicked strand
- Phosphate has good LG
- Base catalyzed 3’ OH attack seals strand
Single stranded binding proteins
Keep ssDNA from reannealing
Telomere
Sequence of repetitive sequences at the ends of chromosomes
Telomerase
Carries RNA fragment
Template DNA synthesis at end of telomere on parental strand
Prevent shortening of strands
Main pathways to mutation
Proofreading missed a mistake
Modified base leads to incorrect pairing
Distinguish parent from daughter strand for repair
Mark parental strand with methylation marks
Mismatch Repair Pathway
Mismatch detected b MutL-MutS
DNA threaded through complex until encounter of MutH
Complex cleaves unmodified daughter strand
Nuclease and helicases degrade unmethylated strand
Gap filled in by DNA Pol III
Sealed by ligase
Alkylation
Add methyl or ethyl groups to nucleobase
Deamination
Replace amine with carbonyl group on nucleobase
Thymine is immune
Depurination
Loss of entire purine base from nucleotide