chapter 11 part 3 Flashcards
when do we do non-homologous end joining?
when there is a DSB that occurs during G1 (before replication) which prevents completion of DNA replication
what does non-homologous end joining allow for?
cells to regain the ability to complete DNA replication, though we will have mutations for sure
non-homologous end joining process
4 steps
-DSB happens
-DSB are recognized by the proteins PKcs, Ku70, and Ku80 that attach to the broken ends of the DNA
-the complex trims back the free ends of the break- LOSS OF GENETIC INFO
-the blunt ends produced by resection are ligates by ligase IV
when does synthesis-dependent strand annealing SDSA occur?
we do it when there are DSBs that occur after replication by an error-free process
what is synthesis-dependent strand annealing also called?
homology dependent repair (HDR)
what does the strand invasion process do?
displaces one strand of the DNA duplex on the sister chromatid, forming a displacement (D) loop
-replication within the loop synthesizes new DNA from the intact template strand
what is the process of SDSA
-SDSA begins with trimming the broken ends, followed by attachment of protein Rad51 (a homolog of bacterial RecA protein)
-Rad51 facilitates the invasion of the intact sister chromatid by the resected end of the broken strand
-stand invasion process
-sister chromatids refrom
how do sister chromatids reform in SDSA?
by dissociation & annealing of the new strand to the other side of the break, resulting in replacement of the excised DNA with a duplex identical to the sister chromatid
homologous recombination
the exchange of genetic info between homologous DNA molecules
homologous recombination in bacteria
occurs during conjugation & as a consequence of DSB repair (archaea as well)
homologous recombination in eukaryotes
takes place in prophase 1 of meiosis
what is homologous recombination initiated by in eukaryotes?
double-strand DNA breaks
what does proper chromosome segregation in meiosis depend on?
homologous recombination
-without it, errors like nondisjunction are likely to occur
what is the system of homologous recombination in bacteria?
RecBCD pathway
what does RecBCD rely on?
DNA double-strand breaks to innitiate the process
-this attracts the RecA protein (homolog of human Rad51 protein)
-RecBCD then attaches to the region where RecA is bound, leading to single-strand invasion & D loop formation (similar to SDSA)
-RuvAB & RuvC proteins bind & complete homologous recombination
DSB model of meiotic recombination simplified
- recombination initiated by Spo11
- Spo11 degrades, Mrx cuts single strand of cut chromatin
- Rad51 & Dmc1 facilitate strand invasion & D loop formation
- holliday junction formed between 2 strands that appear to cross over
- heteroduplex forned
- invading strand in extended with DNA synthesis guided by intact template strands assisted with Rad52 & Rad59
- 3’ end of invading strand joins 5’ end of a strand segment that was initially invading strand (ligation)
- non-sister chromatids are connected by double holliday junctions (DHJs)
heterodulex
a double-stranded DNA formed from single stranded pieces of DNA of different homologs
how many ways can holliday disjunction be resolved
2
what are the way holliday disjunction can be resolved
-same sense resolutions
-opposite sense resolution
same sense resolution
when two north-south (NS) resolution cuts or two east-west (EW) resolution cuts occur
-flanking markers DO NOT recombine though heteroduplex regions remain only in between junction points