Homologous

Recombination
@ Molecular
Level
is a major
mechanism used to
repair DSBs

Double standard breaks [DSBs]

• arise frequently in DNA & if not repair cell is disastrous.
• these broken DNA caused by several mean. - mutation.

[Prokaryote]
HR repair DSBs promotes genetic exchange in bacteria.
• the exchange occurs btw. the chromosome of one cell
& DNA that enter the cell via phage-mediated
transduction @ cell-to-cell conjugation.
• the new DNA [linear molecule] enter the cell & thus
provide the critical ‘broken’ DNA end that needed to
initiate the recombination.
[Eukaryote]
HR is critical for repairing DSBs & collapse replication
fork
• HR is essential to the process of chromosome pairing
during meiosis.
• in this case, as cells enter meiosis,
• a specific protein introduced DSBs into DNA and
initiate the recombination pathway.

Cleavage of Holiday Junction

A. Alignment of two homologous DNA molecules.
• DNA identical region >100bp or so
• DNA can have small regions of sequences different i.e different
sequence variant (alleles)

B. introduction of breaks in the DNA, which then processed to generate

regions of single-stranded DNA (ssDNA).

C. Strand invasion — occur when ssDNA of parental paired with

complementary strand in homologous DNA duplex.
• result- regions of new duplex are generated in which contain some
mismatched basepair (heteroduplex DNA)

D. After invasion, two DNA molecules become connected by a crossing

DNA [called Holiday Junction].
• the branch migration occurs when the identical base pairs are
formed in the recombination intermediate.

• Cleavage of Holiday Junction is DONE by a process called

[Resolution}
• cutting the DNA strand within the holiday junction regenerates two
 Homologous Recombination in Prokaryotes [E.coli]
— protein that promote HP of E.coli via a major DBS-repair pathways [RecBCD pathway]

• the RecBCD enzyme processes broken DNA molecule to generate these regions of sand.
• RecBCD —help load the RecA strand exchange protein onto these ssDNA ends.
• RecBCD composed of :
—3 subunit [products of recB, recC & reD genes
—& DNA helicase & nuclease activiities
• it bind to DNA molecules at the site of DSB & tracks along DNA using the energy od ATP
hydrolysis.
• thus, the DNA is unwound, with or without the accompanying nucleolytic destruction of on or
both of the DNA strands.
• the actives of RecBCD is controlled by a specific DNA sequence - chi sites.

Homologous Recombination in Eukaryotes

— is most critical for meiosis.

• During meiosis, HR is required for :

• proper chromosome pairing & thus maintaining the integrity of the genome.
• HR also reshuffle genes between parental chromosome, ensuring variation in the sets of genes
passed to the next generation.

Homologous Recombination ore required for chromosome segregation during meiosis.

• meiosis involves two round of cell division, resulting in reduction of the DNA content from
diploid(2n) to haploid(n).
• before cell division, the cell has two copies of each chromosome (homologs), one each that was
inherited from its two parents.
• During S phase, these chromosomes are replicated to give a total DNA content of 4N.
• the products of replications — that is , the sister chromatids-stay together.
•
• then, in preparation of the first nuclear division, these duplicated homologous chromosomes
must pair & align at the center of the cell.
• it is this pairing of homologs that requires homologous recombination

• Then, in second nuclear division, it is sisters chromatids that separate.

• The products of this division are four gametes (1N each chromosome)
• the homologous recombination events that occur during meiosis are called {meiotic recombination]

• in the absence of HR, chromosome often fail to align properly for the first meiotic division.
• results : high incidence of chromosome loss
• improper segregation of chromosomes [nondisjunction]
• leads to large number of gametes without correct chromosome complement [poor fertility]
• Meiotic recombination
• frequently give rise to crossing over btw genes on the two homologous parental chromosome
GENETIC CONSEQUENCE OF HR

Gene Conversion — observed during normal HR event, i.e in genetic exchange in

bacteria & pairing chromosome during meiosis.
Two ways of gene conversion occurrence:
1. the 3’ sand tails invade the homologous duplexes and elongated, cause the copy
of ‘a’ information could replace ‘A’ information in the products chromosome upon
completion of recombination

2. the repair of base pair mismatches that occur in the recombination intermediate.