Recombination 1 Flashcards
What is recombination?
- Allows favorable and unfavorable mutations to be separated and tested as individual units in new assortments
- Provides a means of escape and spreading for favorable alleles and a means to eliminate an unfavorable allele without bringing down all the other genes it is associated with
- Basis of natural selection
when does recombination occur?
During gamete formation
- Two sister chromosomes divide forming chromatids
- 4 chromatids
- The crossing over occurs with the chromatids, but only with 2 out of the 4
- 50% recombinants
homologous recombination
- Homologous (generalized): in eukaryotes, it occurs at meiosis in males and females in the formation of the gametes
- Takes 50 bases being identical before this occurs in bacteria
- Occurs during meiosis
- Does not change the order of the genes - no change in genome organization
site-specific recombination
phage integration; inversion of specific regions of the bacterial chromosome
- Site specific recombination changes the genome organization = change order of genes/configuration; deleted or switch order
what are some examples of site-specific recombination?
- VJ joining in mammalian antibody genes
- mating type switch
leptotene
- chromosomes first visible, DNA replication has already finished
- condensed chromosomes become visible, often attached to nuclear envelope
zygotene
- chromosomal pairing begins
- chromosomes begin pairing in limited region/s
- pair by the pattern of proteins on the DNA
pachytene
- synaptonemal complex complete; recombination is completed
- Synaptonemal complex extends along entire length of pair chromosomes
- Synapsis is chromosomal pairing
- Thick complex
diplotene
- chiasmata are visible
- Chromosomes segregated but are held together by chiasmata
- Chiasmata - formation of chi (x formation)
diakinesis
- chiasmata are visible
- Chromosomes condense, detach from envelope, chiasmata remain, all 4 chromatids become visible
what are the steps of homologous recombination?
- Homologous recombination occurs between synapse chromosomes of the gametes
- Chromosomes must synapse (pair) in order for recombinant nodes to form where crossing over is thought to occur
- Leptotene: chromosomes first visible; DNA replication has already finished
- Zygotene: chromosomal pairing begins (synapsis)
- Pachytene: synaptonemal complex complete; recombination is completed
- Diplotene: chiasmata are visible, chromosomes separated
- Diakinesis: chiasmata are visible, chromosomes condense and all 4 chromatids are visible
when do ds breaks begin?
- Leptotene
- Homologous recombination always starts at one of the two chromatids
When is recombination complete?
Late pachytene
what are the 5 ways to repair a double stranded break in eukaryotes?
- double-strand break repair
- synthesis-dependent strand annealing
- nonhomologous end joining
- single strand annealing
- chromosome break induced replication
double-strand break repair
- ds break in one pair of the DNA mediated by GEN1
- 5- end resection - nuclease chews the 5’ end on either side of the break and happens immediately after break is recognized
- single strand invasion of intact DNA - the 3’ end produced a single strand with a 3’, protruding into the second set of DNA - this serves as a primer for replication/synthesis
- synthesis of both strands - single strand invasion into the intact DNA forms a D-loop duplex
- branch migration expands the D-loop, has to expand past the 5’ end is in order to trigger end capture
- when the ends join together is called end capture
- need homology to form the second holiday structure
- double holliday structure forms
- When you get the displacement loop, it joins with the top molecule (anneal) so you have to have enough exposed to form with the upper strand to repair the break at the top as well
- ligation occurs
- outcome is cutting which can result in DSB crossover or no crossover
when does DSBR occur?
- damage
- collapsed replication fork
branch migration
- branch sites can migrate in either direction
- this is an important step in expanding the D-loop to reach the far side of the gap left by the ds break
- movement probably driven by the recombination machinary
when is the DSBR pathway used?
- ds breaks during accidental breaks during mitosis, outcome is no crossover
- introduced ds breaks during meiosis, outcome is both crossover or no crossover, with a heavy bias towards crossover