Q2-FUN10/DNA Replication Flashcards
why is eukaryotic DNA replication so difficult?
there is a large amt of DNA to be replicated, chromosomes are strucutrally complex, need a lot of enzymes, takes a lot of time
how long is the cell cycle for yeast
1.4 hrs
how long is the cell cycle for cultured animal cells?
16-24 hrs
how long is the cell cycle for human cells
8hrs - 100 days or permanent (G0)
what does semiconservative replication mean?
two copies of the original dna molecule are produced, and each copy replicates the info from 1/2 of the original DNA molecule
what are the 4 DNA replication requirements?
- single stranded template
- nucleotides
- replisome
- a primer with a free 3’ hydroxyl group
what is a replisome?
large protein complex that carries out DNA replication, has a bunch of enzymes and proteins in it
what is the origin of replication?
a place where the separation of 2 complimentary strands occur
what moves outwards from the ori during initiation?
two replication forks
what are the unwinding proteins?
DNA helicase, single-strand binding proteins, and topoisomerase
what does DNA helicase do?
it separates the DNA strands, uses energy
what do SSB proteins do?
these proteins bind single stranded regions of DNA to prevent the strands from re-associating
what does topoisomerase do?
it regulates the twisting of DNA, prevents DNA supercoiling
What is primase?
it is a RNA polymerase enzyme
What does primase make?
it makes a primer, which is an RNA starter strand
the primer starts on the __’ to ___’ strand
5’ - 3’
Why do we need RNA polymerase to make a primer?
because DNA polymerase enzymes can only extend a chain, they cant make DNA from scratch
they require a free 3’ hydroxyl group to extend DNA (DNA is copied in the direction of 5’ to 3’)
once RNA primer is removed, what fills its gap?
DNA polymerase
what does DNA poly use as a template?
single stranded DNA
how does DNA poly read its template?
3’ to 5’
how does DNA poly MAKE new DNA?
5’ to 3’
DNA starts the formation of __________ bonds
phosphodiester
What does PCNA stand for?
proliferating cell nuclear antigen
what does PCNA do?
it acts as a sliding clamp. DNA polymerase attached nucleotides at abt 1000 per second so it needs something to keep it close to the template as it moves along.
What enzyme helps proofread the activity so that no errors occur?
DNA polymerase
what is substrate specificity in DNA replication?
the DNA poly active site can bind all 4 nucleotides, but catalysis only occurs when the correct one is bound
what is 3’ to 5’ exonuclease activity?
it is the proofreading of the strand. it removed nucleotides at the 3’ end of the new strand that is incorrectly matched.
which strand is made continuously and which strand is made discontinuously?
leading, lagging (DNA poly can only make DNA 5’ to 3’)