DNA RECOMBINATION

once a foreign piece of DNA has entered a cell through one of the gene exchange systems what
happens to it? if the host genome and the new DNA have a considerable stretch of homologous
DNA sequence in common that is the sequences are similar, but not necessarily identical then the
foreign DNA may be incorporated into the chromosome through a process called generalized
recombination.
enzymes participating in generalized recombination find and align homologous stretches of DNA
in two different DNA molecules and catalyze an exchange of strands. an enzyme system called the
recBCD complex, initiate the series of events by binding to the end of a DNA fragment and
beginning to unwind it.
a specific eight base sequence, called a chi site, exists periodically in most DNA sequences and
serves as a recombinational hotspot. recBCD unwinds the DNA until it reaches such a site.The
complex then mix the DNA and continues unwinding as it loads a protein called RecA onto the 3-
prime tail of the single-stranded DNA. the recA covered DNA strand is called a recA filament.
RecA molecules also called synaptases can scan DNA molecules for homology and align the
homologous regions. this reaction requires a free 3’ prime end on the single-stranded molecule and
results in a triplex DNA molecule called a synapse.
notice that the strand invasion reaction results in a deal loop formation (shaped like the letter D).
the donor single-stranded DNA has invaded the homologous region in the double-stranded DNA
recipient molecule and displaces it's like strand.
at this point a single strand crossover called a Holliday Junction has been made.
Other recombination proteins called RuvA and B, assemble at the crossover point and pull the
donor and recipient DNA strands in opposite directions, extending the invasion, in a process called
strand assimilation or branch migration. this process extends the base pairing between homologous
donor and recipient strands.
ultimately the end of the displaced strand at the D loop is cleaved and the donor strand is ligated
to the recipient strand.
rotating one of the molecules at the branch point provides more clarity for the events that occur
next and reveals a classic holiday structure.
an enzyme called RuvC can cleave across the junction in either of two ways. in one case, cleavage
and ligation of two of the strands results in a closed circular molecule and a linear molecule, similar
to the starting molecules except that a DNA segment has been exchanged between them.
in the second cleavage option, cleavage and ligation of two of the strands results in a linear
molecule. the loss of the chromosomes circularity would result in death for the bacterium.
However, a second crossover event would restore circularity to the chromosome and leave it with
a swapped segment of DNA.
let's look again at the second cleavage option. this time imagine the original donor molecule not
as a linear fragment, but as circular. in this case, the result of recombination is a cointegrate
molecule, where a single crossover joins the two participating DNA molecules into a larger circular
Molecule.
what can generalized recombination due for a microbial cell? cells with damaged chromosomes
may use donated DNA to repair their damaged genes. recombination may also be part of a “self-
improvement” program that samples genes from other organisms to enhance cellular fitness.
recombination also provides an internal method of DNA repair, useful to fix mutations or restart
stalled replication forks. However, this last role does not involve foreign DNA