Lippincott - DNA Structure, Replication, Repair Flashcards
DNA
polymer of deoxyribonucleoside monophosphates covalently linked by 3’–>5’ phosphodiester bonds
main bond in DNA
phosphodiester bond
phosphodiester bond connects:
3’-hydroxyl group of one nucleotide to 5’ hydroxyl group of adjacent nucleotide
3’ end
end with the free hydroxyl
5’ end
end with the free phosphate
what enzyme family cleaves DNA
deoxyribonucleases
what enzyme family cleaves RNA
ribonucleases
orientation of single stranded DNA in double helix
antiparallel
type of bond that mediates base pairing
hydrogen bond
DNA - adenine pairs with:
thymine
DNA - guanine pairs with:
cytosine
DNA - thymine pairs with:
adenine
DNA - cytosine pairs with:
guanine
methods of DNA denaturation
altering pH (ionization), melting DNA
number of H bonds between guanine and cytosine
3
number of H bonds between adenine and thymine
2
which bases are purines
adenine, guanine
which bases are pyrimidines
thymine, cytosine
number of rings in purines
2
number of rings in pyrimidines
1
what base pairing denatures at higher temperatures? why?
GC, because of greater number of H bonds
process of reforming the double helix from single strands of DNA
renaturation or reannealing
process of separating the double helix of DNA
denaturation
major stuctural forms of the double helix
B form, A form, Z form
B form
right handed helix with 10 residues per turn, main form of chromosomal DNA
A form
right handed helix with 11 residues per turn, produced by dehydrating the B form
Z form
left handed helix with 12 residues per turn, occur naturally in regions with alternating purines and pyrimidines (ex. poly GC)
region where active DNA synthesis occurs
replication fork
DnaA protein
binds to specific nucleotide sequences at origin, causing AT rich regions in the origin to melt
DNA helicase
actively unwinds the double helix
SSB
single strand binding proteins; keep the ssDNA separated and protect it from nucleases
type of enzyme that counteracts supercoiling
DNA topoisomerases
DNA topoisomerase I
relax both negative and positive supercoils, does not use up ATP
DNA topoisomease II
relaxes both negative and positive supercoils, requires ATP
DNA gyrase
creates negative supercoils in order to counteract positive supercoils
enzyme that copies the DNA templates
DNA polymerase
DNA polymerase - direction of reading
3’–>5’
leading strand
3’–>5’ strand of template; needs only one promoter, synthesized continuously
lagging strand
5’–>3’ strand of template; needs multiple promoters; synthesized discontinuously
stretches of discontinuous DNA in lagging strand
okazaki fragments
RNA primer
short stretch of RNA required by DNA polymerase to begin replication
responsible for RNA primer synthesis
primase
responsible for catalysis of DNA chain elongation
DNA polymerase III
DNA polymerase - direction of synthesis
5’–>3’
substrates of DNA polymerase III
5’ deoxyribonucleoside triphosphates
when each new deoxynucleoside monophosphate is attached to the growing chain by DNA polymerase III, what is released?
pyrophosphate
for DNA synthesis to occur, what 4 substrates must be present?
dATP, dTTP, dCTP, dGTP
2 main activities of DNA polymerase III
5’–>3’ polymerase; 3’–>5’ exonuclease
5’–>3’ polymerase activity of DNA polymerase III
for synthesis of new DNA strand
3’–>5’ exonuclease activity of DNA polymerase III
for proofreading
responsible for excision of RNA primer
DNA polymerase I
activities of DNA polymerase I that allows it to complete its function
[1] 5’–>3’ exonuclease activity to remove the primer
[2] 5’–>3’ polymerase activity to fill in the gap with deoxyribonucleotides
[3] 3’–>5’ exonuclease acitivity to proofread
difference between 5’–>3’ exonucleases and 3’–>5’ exonucleases
5’–>3’ exonucleases can remove groups of altered nucleotides (up to 10) in the 5’–>3’ direction
exonucleases used by DNA polymerase III
3’–>5’ exonuclease
exonucleases used by DNA polymerase II
3’–>5’ exonuclease
exonucleases used by DNA polymerase I
3’–>5’ exonuclease, 5’–>3’ exonuclease
responsible for sealing nicks between okazaki fragments after excision and replacement of RNA primers
DNA ligase
DNA polymerases in prokaryote replication
DNA polymerases I, II, III
DNA polymerases in eukaryote replication
DNA polymerases alpha, epsilon, delta, beta, gamma
DNA polymerase alpha
has primase activity
DNA polymerase beta
for gap filling in DNA repair
DNA polymerase gamma
replicates mitochondrial DNA
DNA polymerase delta
elongates okazaki fragments in lagging strand
DNA polymerase epsilon
elongates leading strand
telomeres - structure
complexes of noncoding DNA + proteins at ends of chromsoomes
telomeres - function
mediate structural integrity, protect from nuclease attack, allow repair systems to distinguish ends of DNA
responsible for maintaining telomeric length
telomerase
reverse transcriptase
synthesize DNA from RNA template
classes of histones
H1, H2A, H2B, H3, H4
charge of histones at physiologic pH
+
make up core of histone beads
H2A, H2B, H3, H4
location and function of H1
bind to linker DNA between beads, facilitates packing of histones into more complex structures
effect of UV radiation on DNA
fusing together of pyrimidine to form pyrimidine dimers (ex. thymine dimers)
effect of high energy ionizing radiation on DNA
double strand breaks
principle of methyl-directed mismatch repair
recognition of the parent strand as the methylated strand –> repairs are then based on the parent strand
responsible for recognizing mismatch in methyl directed mismatch repair
Mut
enzyme involved in repair of damage from UV radiation
UV specific endonuclease
xeroderma pigmentosum
inability to repair UV damage, extensive mutation due to exposure to sunlight
involved in removal of abnormal bases
specific glycosylases
involved in recognition and repair of AP site
AP endonucleases (recognition of missing base), deoxyribose phosphate lyase (removal of base-free sugar phosphate), DNA polymerase, DNA ligase
deoxyribose phosphate lyase
removes the base-free sugar phosphate residue in repair of AP sites
methods of repair of double strand breaks
[1] nonhomologous end-joining repair
[2] homologous recombination repair
nonhomologous end-joining repair
crude ligation of DNA on either end of double strand break; prone to mutation (some DNA is lost)
homologous recombination repair
use of homologous DNA as a template to restore lost DNA from double strand breaks
enzymes in homologous recombination repair
enzymes that perform genetic recombination between homologous chromosomes during meiosis