Genetic mechs Flashcards
What did Levene correctly and incorrectly determine in DNA structure
DNA rep history
Corr - DNA made from nucltid, structure of nucleotide
Incorr - Tetranucleotide hypothesis (bases form circle, equal ratio of all bases)
What did Chargaff figure out and how did it refute Tetranucleotide hypothesis (levene)
DNA rep history
A pairs with T
G pairs with C
Ratio of A and T does not affect G, C
Ratios can be diff
What was error in pauling triple helix model
DNA rep history
not triple helix
phosphates were located in core of helix
negative O would repel in reality
what was technique used by franklin and wilkins to determine DNA structure
DNA rep history
X ray crystallography - DNA sample crystallized, shoot xrays, measure defraction to find 3d structure
signifigance of watson and cricks base pairing
DNA rep history
finalized structure
inspired by alpha helix protein shape
won prize
found location and angle
conservative vs semi conservative vs dispersive model
DNA rep
conservative - whole parent DNA preserved, complete new copy made
semi cons - correct - half parent, half new in each
dispersive - bits of parent, bits of new in chunks
What were the steps in Meselson and Stahls exp
DNA rep
- grew ecoli in heavy nitrogen (N15)
- transfer ecoli to lighter N14, 1 reproduction, centrifuge
- see 1 band (cant be conservative )
- 1 more replication in N14, centrifiuged
- see 2 bands (cant be dispersive)
Must be semi conservative
What would each DNA rep model look like after the first centrifuge
DNA rep
conservative - 2 band (complete parent , complete new)
semi cons- 1 (2 half/half)
dispersive - 1 (mix of both averages out)
What would each DNA rep model look like after the second centrifuge
DNA rep
cons - 2 (3 new on top, 1 parent bottom)
semi cons - 2 (2 half/half, 2 fully new )
disruptive - 1 (average density)
difference between prokaryote and euk DNA rep init
DNA rep
prok - rep begins at 1 certain point, 1 bubble expands
Euk - rep begins thousands of places at once on dna, bubbles form
What is an Ori
DNA rep
Origin of replication
Special sites on DNA where replication begins
Contains a specific sequence recognized by the replication machinery (enzymes)
Often high in A-T base pairs
Steps and proteins in DNA rep init
DNA rep
initiator protein unwinds the DNA
helicase seperates strands
Single-strand binding proteins (SSBPs) - keeps them apart
Topoisomerase
primase makes primer
Why is priming needed
DNA rep
New nucleotides can only be added to the 3 end
DNAP I vs DNAP III
DNA rep
both found in prok
DNAP III catalyzes elongation, adds nucleotide to existing 3’ end, ntide is hydrolyzed
DNAP I - replaces the RNA primer with DNA
what is the problem wtih rep fork as it relates to anti parallel nature of DNA
DNA rep
due to anti parallel nature of DNA one of the new DNA strand will have 3’ end at fork and the other will have 5’ end at fork
New nucleotides can only be added to 3’ end
but both new DNA strands must be built
diff in steps and enz for leading and lagging strand
DNA rep
leading - primase makes 1 primer,DNAP III builds off it, 1 DNAP used at ori
lagging - primase makes primer, DNAP III builds off it, DNAP I replaces primer, DNA ligase, happens multiple times
what is okazaki fragment
DNA rep
fragments of new DNA on lagging strand
What is leading and lagging strand
leading - synthesized continuously, polymerized in direction of fork
Lagging - synthed in short okazaki fragments, polymerized opposite direction of fork
what is telomere
DNA rep
bunch of useless DNA at end of chromosome
starts with 15000 at cenception
death with 5000
What does topoisomerase do
DNA rep
breaks DNA bonds and reforms them
helps to unwind DNA and lower tension
on either side of bubble
What does helicase do
DNA rep
disrpts H bonds in DNA and seperates DNA to make the fork
in base of fork
What do single stranded binding proteins do (SSBPs)
DNA rep
binds to unwound DNA strands keeps it apart
What is an RNA primer
DNA rep
made by primase so that the DNA has something to attach to
is eventually taken out
WHat is 3’ and 5’ end
DNA rep
Has to to with which carbon is exposed.
Nucleotides can only be added to the 3’ end.
What does DNAP I do
DNA rep
replaces the RNA primers with DNA, takes place after the DNAPIII
if DNA nucleotides can only be added to a 3’ end, how does DNAP I add new DNA?
it adds it to the 3’ end of the strand before it
What does DNAPIII do
DNA rep
Adds to the 3’ end of an existing strand
What does DNA ligase do
DNA rep
Binds together the okazaki fragments after they are completely DNA
also used to bind the 2 DNA at ori
uses phosphodiester bonds
Eukaryote vs Prok rep termination
DNA rep
euk - when bubble reach another bubble, when it reaches end of DNA
prok - when it circels around to beginning of bubble
what is the replication problem at the end of linear DNA
DNA rep
There was a primer used to start to make the 5’ end that was removed meaning that the ends do not line up, some of the original DNA was not copied. happens with the lagging strand
what is purpose of telomeres and structure
DNA rep
found at end of euk chromosomes
noncoding
made from repeats of short genetic sequence
protects DNA from being eroded, getting unound
what is senescence
DNA rep
when the cell stops growing or dividing, recognizes that DNA is damaged, will not divide for protection
what is apoptosis
DNA rep
programmed self descructtion
what is the cellular concequence of shortened telomeres
DNA rep
senescence or aptosis
what is telomerase and what does it do
DNA rep
enzyme that adds telomeres to chromosomes
is a ribonucleoprotein
does reverse transcriptase
what is a ribonucleoprotein
DNA rep
protein that contains RNA in it and protein
ex human telomerase contain RNA (AAUCC)
what is reverse transcriptase
DNA rep
When RNA is used to make DNA instead of the other way around
ex. Telomerase uses its internal RNA to make the DNA for the 3’ of chromosmes (makes the longer part longer )
How do telomeres relate to aging
DNA rep
they get shorter as you age
15000 bases at conception
10 000 at birth
5000 at death
How the telomere is completed using telomerase
DNA rep
- binds to 3’ (parent strand)
- extends the parent using its RNA template
- the other complimetnary strand gets primer with primase
- DNAP alpha is used to elongate
- DNA ligase connects the newcomplementary strand with the rest of the complimentary strand
- still end up with the parent strand being longer than the complimentary strand but technically nothing is lost. (eveything needed is copied)
What cells contain telomerase acivity
DNA rep
germ cells and cancer cells
Do eukaryotes use DNAP I and III
DNA rep
No
eukaryotes use ones named with greek letters (15)
prok have 5 DNAP (roman numerals )
DNAP alpha is like DNAP III but for euk
DNAP also seems to make the primer?
DNAP delta is like DNAP I?
why does DNAP alpha make telomeres instead of DNAP III
DNA rep
Telomeres only exist in eukaryotes
the issue only needs to be solved in eukaryotes
DNAP III is used in prokaryotes only and does not need to be able to make telomeres
DNAP alpha is used in eukaryotes only
What is a gene
Transcript
stretch of DNA that is transcribed into RNA
aka transcription unit
What is a promoter and why does it have rich AT regions
Transcript
Where the RNAP first binds
upstream of gene
holds TATA box
holds transcription start point
Rich in AT because they are easier to break apart and start transcription
transcrition initiation in euk vs prok
Transcript
prok - RNAP recognizes promoter, binds, unwinds DNA, starts
euk - pre - initiation complex made (protein factors + anactive RNAP II), RNAP gets phosphorylated at C terminal domain, starts
What is coding strand
Transcript
strand of DNA whos base sequence is identical to the RNA
not the template
What is the sense strand
Transcript
same as the coding strand
it is the one what looks like the RNA
what is the antisense strand
Transcript
the template for the RNA
complimentary to the sense strand
does not mean anything by itself
What is the RNA Transcript
Transcript
WHat initially comes out of transcription before the post processing
primary Transcript
newly synthed RNA
Where does Transcript take place
Transcript
euk - nucleus
prok - cytoplasm
How do you know the 3’ end 5’ of DNA based off RNA
Transcript
RNA must be made be made in direction of 3’ end
sense strand is in same direction as RNA
antisense strand / templete - is opp direct of RNA
Transcription termination for euk
Transcript
- dephosphorylation of C terminal domain on RNAP II (slows it down)
- AAUAAA gets Transcribed, stalls RNAP II, but continues
- GU rich region, gets transcribed, signals endonuclease to cut
- cut happens after AAUAAA, before GU region on RNA
transcription termination for prok
Transcript
- 2 GC rich regions that are complimentary
- when transcribed form hairpint
- dA region on DNA make U region on RNA, weak bonds
- RNAP stops and is released
How are Transcription termination different euk vs prok
Transcript
prok - uses hairpin method
Euk - uses endonuclease to cut RNA off from RNAP
What are the types of RNAPs used euk vs prok
Transcript
prok - only hase 1 types of RNAP
Euk has 3 types, mainl RNAP II used
What is mRNA
Transcript
messenger RNA
final transcript, what gets translated
leaves neucleus
What are the 3 types of RNA processing do both euk and prok have it
Transcript
capping
polydenylation
splicing
only euk does this
What is capping (what, where, why, when)
Transcript
at 5’ end of new RNA
addition of modified guanine
protection from degredation, helps ribosomes bind
during transription