The Role of RecA in DNA Replication

Hazem Dawoud

http://www.callutheran.edu/BioDev/omm/reca/recamast.htm
 Outline

* introduction.
* RecA binding sites.
* RecA functions and activities.
* SOS response.
* TLS
* RecA filament formation on DNA.
* Repair of a collapsed replication fork by RecA protein.
DNA replication
DNA Replication
RecA protein :
38 kilodalton polypeptide.

identified in E. coli by Clark and

Margulies in 1965.

DNA repair and maintenance.

Class of homologous DNA repair protein.

More than one DNA binding site.

DNA dependent ATPase : allosteric enzyme.

RecA Binding sites:

DNA binding sites.

ATP binding sites

Nature Reviews Molecular Cell Biology 8, 127-138 (February 2007) |

RecA protein functions:

Repair of stalled replication fork

double-strand break repair

general recombination

induction of the SOS response

SOS mutagenesis

Promotes mutagenic Trans-

Lesion Synthesis by polV.
Annu Rev Microbiol. 2003;57:551-77. Cox MM.
 Activities of Escherichia Coli RecA:

a-Strand invasion
reaction d-The replication
fork regression
reaction
b- a three-strand-
exchange reaction e- the RecA
coprotease function.

C-a four-strand-
exchange reaction. f- stimulation of
translesion -DNA-
replication activity
of DNA pol 5

Nature Reviews Molecular Cell Biology 8, 127-138 (February 2007) |

 The SOS Respose:

In non-induced growth conditions, the LexA

repressor binds to
SOS-controlled promoters, limiting or
preventing their action.

RecA filaments formed at sites of damage

activate the autocleavage of the LexA
repressor, allowing SOS gene expression.

SOS induction is reversed when damages

are repaired.

PLoS Biol. 2005 July; 3(7): e255. Bénédicte Michel

TLS :

Trans-Lesion SynthesisDNA damage tolerance

process

PCNA, proliferating cell nuclear antigen,

provides a platform for TLS Pol.
Involves switching out regular DNA pol with
specialized trans-lesion pol.
Low fidelity on the undamaged template.
recombination repair
Assembly of RecA filaments:

Cox Nature Reviews Molecular Cell Biology 8, 127–138 (February 2007) | doi:10.1038/nrm2099

01/09/12
 RecA protein Filament Formation on DNA.

RecA protein filaments

are generally formed in two
distinct phases, nucleation
and extension.

Extension proceeds largely in

the 5 to 3 direction, and
cancontinue readily into any
adjacent duplex DNA.

Critical Reviews in Biochemistry and Molecular Biology, 42:41–63, 2007, Michael M. Cox
 Repair of a collapsed
replication fork.

RecA bind to ss DNA and

catalyze hyperdization.

RecA catalyze invasion of

duplex DNA.
 Summary
* RecA protein is DNA binding protein with multiple activiries.

* RecA protein polymerize to form nucleoprotein filament.

*RecA protein promotes mutagenic TLS by pol V.

* RecA protein induces the SOS respose.

* RecA promotes strand invasion to initiate recombination.

* RecA protein repair of stalled replication fork.

 Thank You

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