CH4 DNA, RNA, Flow of Genetic Info P2 Flashcards
During which phases must the 2 strands of DNA double helix be sep’d?
replication
transcription
Denaturation (Melting)
when the double helix is reversibly melted.
DNA strand separation via heating soln.
Hypocrhomism
how DNA melting is observed.
bases stacked in double helix absorb less UV light than those in single-stranded molecule.
Reannealing
process when cooling two strands bind to e/o, reform double helix.
Polymerase
replicate DNA.
catalyze phosphodiester-bridge formation.
instructions from templates.
(DNA)n + dNTP (DNA)n+1 + PPi
DNA replication.
rxn catalyzed by DNA polymerase.
What are the key characteristics of DNA synthesis?
- 4 Deoxynucleoside triphosphates and Mg2+
- template strand
- primer
- DNA polymerase w/ nuclease activity
Deoxynucleoside Triphosphates
activate precursors
Template Strand
directs DNA synthesis
Primer
what the new strand grows from
Nuclease Activity (of DNA polymerase)
allows for removal of mismatched bases
The polymerization reaction is catalyzed by?
DNA polymerases
dNTPs
incorporates a phosphate
activated precursors
subsequent breakdown of released pyrophosphate helps drive phosphodiester bond formation
Pyrophosphate Hydrolysis
drive strand elongation
Tobacco Mosaic Virus
RNA genomes replicated by RNA-directed RNA polymerases
Retroviruses (ex HIV-1)
single-stranded RNA genomes converted to DNA double helices via reverse transcriptase
Flow of Info
RNA to DNA in Retroviruses
Each step uses Reverse Transcriptase
- Viral DNA
- Synthesis of DNA complementary to RNA - DNA-RNA Hybrid
- RNA digestion - DNA Transcript of Viral RNA
- Synthesis of 2nd Strand of DNA - Double-Helical Viral DNA
Reverse Transcriptase
enzyme that converts RNA genome of retrovirus into DNA.
catalyzes synthesis of complementary DNA strand, digests RNA, subsequent synthesis of DNA strand.
Gene Expression
transformation of DNA Info into functional molecules
What are the most abundant types of RNA?
tRNA
mRNA
rRNA
RNA polymerase
catalyzes transcription (synthesis of RNA from DNA template)
initiates and elongates RNA product w/ chain growing 3’ to 5’
no primer needed
3’ OH of growing chain attacks innermost phosphoryl alpha group of incoming nucleoside triphosphate
What are the 3 requirements of RNA polymerase?
- DNA template
- activated precursors in form of 4 ribonucleoside triphosphates
- Divalent metal ions (ex Mg2+, Mn2+)
DNA Template
complementary seq of newly synthesized RNA
Coding Strand
DNA strand w/ same seq as RNA product (w/ T instead of U)
RNA polymerase catalyzes this rxn:
(RNA)n residues + ribonucleoside triphosphate (RNA)n+residues + PPi
Transcription Mech of Chain-Elongation Rxn Catalyzed by RNA Polymerase
driven thermodynamically by hydrolysis of pyrophosphate
Transcription begins near?
promoter sites
Transcription Ends at?
terminator sites
Promoters
specific DNA seq’s that direct polymerase to proper initiation site.
Consensus Seq
average variation in seq of promoter for diff genes
Promoter Sites for Transcription in Prokaryotes
-35 Region
-10 Pribbenow box
+1 Start of RNA
Promoter Sites for Transcription in Eukaryotes
-75 CAAT box
-25 TATA box
+1 Start of RNA
Elongation continues until?
detects termination signal
Transcription Termination 1: Palindromic DNA
simplest stop signal is a transcribed product of a segment of this
Transcription Termination 2: Hairpin Loop
RNA complement of DNA stop signal forms this followed by several uracil residues
Transcription Termination 3:
Hairpin Synthesis
polymerase stalls
RNA product released
DNA double helix reforms
Transcription Termination 4:
Protein
rho
sometimes req’d
Post Transcription (Eukaryotes): mRNA Modification
5’ nucleotide “cap” added
3’ poly(A) tail added
What are the adaptor molecules in protein synthesis?
tRNA
tRNA
react w/ specific AAs in rxn catalyzed by aminoacyl-tRNA synthetases
Anticodon
template recognition site in tRNA molecules
consists of 3 bases
recognizes a complementary codon
Codon
complementary 3 base seq in mRNA
Aminoacyl-tRNA
attached at 3’ end of RNA
anticodon is template-recognition site
cloverleaf structure w/ many H-bonds btw bases
Protein Synthesis is a process of?
translation
NA seq info translated into AA seq info
The genetic code links?
NA info and AA info
Genetic Code Characteristics
- 3 nucleotides (codon) encode AA
- nonoverlapping
- no punctuation
- has directionality, read 5’ mRNA to 3’
- degenerate - some AA’s encoded by >1 codon, minimizes deleterious effects of mutations
mRNA
translated on ribosomes
1st codon almost always AUG - codes for methionine
AUG in prokaryotes
preceded by purine-rich seq Shine-Dalgarno seq.
AUG in eukaryotes
5’ end is initiator codon
The location of the initiator codon establishes the?
reading frame
Start Codons
req’d for initiation of protein synthesis in prokaryotes and eukaryotes
The genetic code is nearly?
universal
Exons
coding regions in eukaryotic genes that are discontinuous.
Introns
noncoding regions
initially detected by e- microscopy
size range 50-10,000 nucleotides
~ human has 8, some > 100
RNA processing generates?
mature mRNA
Eukaryotic Pre-Messenger RNA
contains exons and introns
1st modified by 5’ cap and 3’ poly(A) tail
mature mRNA generated by splicesosomes
Splicesosomes
large complexes that splice introns to generate mature mRNA
recognize specific seq’s w/i intron that specify splice sites
Introns almost always begin w/?
GU
Introns almost always end w/?
AG
Once mRNA precursors are spliced mature mRNA if formed because these come together.
Exon 1 and Exon 2
TPA Tissue Plasminogen Activator
was generated by Exon shuffling
encodes enzyme that functions in hemostasis
Alternative Splicing
forms set of proteins that are variants of basic motif w/o requiring separate gene for each protein