DNA Mutations,

Damage & Repair

Lecture 5

Cellular DNA Repair Responses

III. Damage Tolerance
SOS Response
Nonhomologous end-joining (NHEJ)

SOS Response

Agents and treatments

UV and X-rays
Alkylating agents
Nalidixic acid
Any treatment that causes halt in DNA replication
SOS phenotypes
Enhanced survival after damage
Enhanced mutation rate
Filamentous growth
Damage inducible genes and function of products
recA
RecA: General recombination; co-protease
lexA
LexA: SOS repressor
uvrABC UvrABC: Exinuclease in Excision repair
umuCD UmuCD: Mutagenesis
sfiA
SfiA: Cell division inhibitor

SOS Response

Consists of three phases:

- The early phase of the SOS response is mostly dominated

by accurate DNA repair.
- The middle phase involves recombination repair.
- The late phase is characterized by elevated mutation
levels caused by error-prone DNA replication
polymerases.

Inducible system that where induction of errorprone replication system is dose dependent

- Higher UV doses induce the system while lower doses do

not.

Regulation dependent on two gene products:

- LexA and RecA

SOS regulation:
Transcriptional regulation via Regulon

Group of genes controlled by one common

regulator - protein repressor LexA

Lex A recognizes a particular operator sequence in

the promoter called the SOS box and binds to it.

SOS Response Regulon is Repressed by

LexA

DNA Damage

Genes controlled by LexA for part of a Regulon

SOS box

Protease Activity of RecA co-protease

activated by DNA damage

LexA is Cleaved by Activated RecA

Induction of SOS Response is Dependent

on Amount of DNA Damage

SOS Response: Relationship between

Induced and Un-induced state
Un-induced State
LexA Repression of SOS Regulon
LexA Repressor Accumulates

Induced State

DNA Damage Signal

High

Low
Normal Repair

Drop in RecA Proteolytic Activity

DNA Damage Signal Reduced

DNA repaired

RecA Proteolytic Activity

LexA Repressor Cleaved

DNA Damage SOS Repair systems Expressed

Error-prone DNA Repair is a Key feature

of the late SOS Response

Error-prone DNA polymerases (umuD operon) are produced only in

the presence of DNA damage via the SOS response.

These specific DNA polymerases attracted to stalled replication forks

due to unrepaired DNA damage and replace normal polymerase.

Sloppy DNA polymerase adds random nucleotides to the strand being

synthesized opposite the damaged bases (template)

Termed Translesion DNA Synthesis

Restores proper base only of the time.

After translesion synthesis, sloppy polymerase replaced by normal

polymerase.

No halt in replication but daughter cells carry new mutations.

Translesion DNA Synthesis

Error-prone Repair

Nonhomologous End-Joining (NHEJ)

Repairs double-stranded breaks (DSB) in DNA

Involves three main proteins (Ku70, Ku80, DNA PKcs) and accessory

proteins

DSB is recognized by Ku dimer (Ku70Ku80) and DNA-PKcs and the

two DNA ends are paired (synapsed).

Other accessory proteins are recruited and DNA-PKcs allows accessory

proteins to process DNA ends

Overhanging nucleotides at end that do not have complimentary

nucleotide to pair are cut back (resect) and ends ligated

Resection results in NHEJ losing DNA error in DNA

Nonhomologous End-Joining (NHEJ)

Resectioning

DNA bases removed mutation introduced