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CHAPTER 24

Recombination

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Figure 24.1
Two Crossovers Result in Recombination
Two paired double-stranded DNA molecules align related regions (A with a; B with b, etc.).
Breakage occurs in each paired DNA molecule followed by crossing over and rejoining of
the ends. This exchanges part of one DNA strand with another.

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Figure 24.2
Homologous versus Non-homologous Recombination
A) In homologous recombination, two DNA molecules have similar sequences such that the pink (top) strand can
align with the purple (bottom) strand. If a double-stranded break occurs within the aligned regions, a crossover
event will exchange regions of the DNA.
B) In non-homologous recombination, related protein recognition sequences lie within two unrelated regions of
DNA. Proteins bind to the recognition sequences and carry out recombination. The proteins direct double-
stranded breakage and crossing over. Genetic exchange can thus occur between two unrelated DNA molecules.
(This event could also theoretically be classified as a translocation.)
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Figure 24.3
Formation of a Crossover
Two homologous molecules of DNA align in regions of similar sequence. A single-stranded
break occurs in the backbone of each molecule. The two ends switch with each other
creating a crossover of single-stranded DNA.

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Figure 24.4
Rearrangement and Resolution of a Holliday Junction
The Holliday junction can isomerize or change into two alternate conformations. Form I isomerizes into the
chi form by rotation of the lower arms at the junction. Another isomerization of the two right arms of the chi
form creates form II. Resolution of the crossover can occur at any time. Resolution of form I exchanges
gene “B” from the pink molecule with gene “b” from the purple molecule resulting in a patch recombinant.
The other strands of the two DNA molecules are unaffected, thus only a small region of heteroduplex DNA
results. For form II, resolution by RuvC hybridizes both DNA strands of the double helix rather than just
one, and both strands are exchanged.
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Figure 24.5
Migration of a Holliday Junction
A complex of four RuvA and six RuvB proteins is able to break and re-form hydrogen bonds
between base pairs, thus allowing the crossover to migrate along the DNA helix.

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Figure 24.6
RecBCD Recognizes Chi Sites
The complex of RecB, RecC, and RecD proteins recognizes the ends of a double-stranded
break and travels along the DNA until reaching the closest chi site. RecD cleaves the
backbone of one strand and dissociates from the complex. RecBC continue to unwind the
DNA beyond the chi site creating a stretch of single-stranded DNA.

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Figure 24.7
RecA Promotes Strand Invasion
RecA binding stabilizes the unwound single-stranded DNA from Figure 24.6. The stabilized
strand is able to invade the homologous double-stranded DNA forming a triple helix.

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Figure 24.8
Integration of Lambda DNA—Overview
Bacterial DNA and l-phage DNA are aligned at the “O” region of the attachment sites. Int
protein induces two double-stranded breaks that are resolved, giving a crossover. Since the
two recognition sites are altered in their flanking regions, l cannot be excised by Int alone
but needs another protein, known as excisionase or Xis, in addition.

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Figure 24.9
Integration of Lambda DNA—Detail of Crossover
The “O” region (green) is cut so that an overhang is generated. Rejoining the cut ends of l
with the bacterial chromosome allows l to integrate its DNA into the host cell.

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Figure 24.10
Timeline of Eukaryotic Recombination in Yeast
Site-specific, double-stranded breaks in DNA appear 60–90 minutes after DNA replication.
The breaks disappear as hybrid molecules are made during the zygotene phase of meiosis.
Resolution of the hybrids then occurs during pachytene. Recombinant molecules appear
approximately 120 minutes after the appearance of double-stranded breaks. Therefore,
eukaryotic recombination occurs in a span of approximately 2 hours.
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Figure 24.11
Spo11 Promotes Double-Strand Breaks
Spo11 binds and cleaves double-stranded DNA in yeast. After cleaving the DNA, Spo11 is
replaced by a nuclease that generates the single-stranded region. The two DNA fragments
are held together by other proteins that are not shown here.

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Figure 24.12
Gene Conversion Following Crossing Over
A) Crossing over between two double-stranded DNA molecules (dsDNA) can occur in a region where two different alleles of the same
gene have a single-nucleotide change. In the blue strand of DNA the “r” allele has the sequence GGCC. In the green strand of DNA,
the “R” allele has the sequence GACC. B) As branch migration of the Holliday junction moves past the nucleotide difference, each DNA
strand now has a mismatched base pair. C) If meiosis occurs before mismatch repair, then the four haploid spores will have two “R”
alleles and two “r” alleles. D) If mismatch repair fixes the mistakes, then one of the mismatched bases will be removed and replaced
with the correct complementary base (shown in red). This Figure shows four possible ways to fix the strands. E) When mismatch repair
fixed the G/T and A/C mismatch, T was changed to C and A was changed to G. After meiosis of the corrected dsDNAs, four “r” haploid
spores are created. F) When mismatch repair fixed the G/T and A/C mismatched base pairs, the G was changed to A and the C was
changed to T. After meiosis, the four haploid spores have only the “R” allele.
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Figure 24.13
Mendelian Ratios in Ascospore Formation
In certain fungi a structure known as an ascus keeps together groups of spores
(“ascospores”) derived from a single zygote. This allows the observation of Mendelian ratios
from a single occurrence of meiosis.

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