ChapLer 27

hage SLraLegles

8acLerlal vlrus
TEM of a T4 phage
Magnification: 450,000 x
27.1 lnLroducuon
!"#$%&'( *$"#+,"-./
01.#'(2
unA repllcauon
SynLhesls of sLrucLural
proLelns (laLe genes)
unA lnLegrauon
no laLe gene
expresslon
lnducuon: sLarL lyuc
cycle
Lyuc cycle and lysogeny are
deLermlned by Lranscrlpuon.
llg. 27.1
27.2 Lyuc uevelopmenL ls
ulvlded lnLo 1wo erlods
A phage lnfecuve cycle ls
dlvlded lnLo Lhe (&"31 perlod
(before repllcauon) and Lhe
3&4( perlod (aer Lhe onseL
of repllcauon).
A phage lnfecuon generaLes
a pool of progeny phage
genomes LhaL repllcaLe and
recomblne.
llgure 27.2
27.4 1wo 1ypes of
8egulaLory LvenLs
ConLrol Lhe Lyuc
5&.6&7(
1. 1wo mechanlsms Lo
recognlze new phage
promoLers:

(1) phage synLheslzes
new slgma facLor
(2) phage synLheslzes
new 8nA polymerase
llgure 27.4
27.4 1wo 1ypes of
8egulaLory LvenLs
ConLrol Lhe Lyuc
5&.6&7(
1. ln
2. lnducuon of
readLhrough by
anuLermlnauon
anuLermlnaLor resulLs
ln fallure ln
Lermlnauon and Lhus
allows Lhe same
promoLer Lo generaLe a
new LranscrlpL.
llgure 27.6
27.3. Lambda (!) lmmedlaLe Larly and uelayed Larly
Cenes are needed for 8oLh Lysogeny and Lhe Lyuc Cycle
Lambda has Lwo !""#$!%&# #%()* +#,#-. / and 0(1, whlch are
Lranscrlbed by hosL 8nA polymerase.
n ls requlred Lo express Lhe $#)%*#$ #%()* +#,#-. lL ls requlred for
boLh lyuc and lysogenlc cycle
1hree of Lhe delayed early genes are regulaLors: 022. 0222. 3
Lysogeny requlres Lhe delayed early genes 022 %,$ 0222.
1he lyuc cycle requlres Lhe lmmedlaLe early gene 0(1 and Lhe
delayed early gene 3.
27.3. Lambda (!)
lmmedlaLe Larly and
uelayed Larly Cenes are
needed for 8oLh
Lysogeny and Lhe Lyuc
Cycle
llgure 27.07
AuLoregulaLory clrculL
llgure 27.08: 1he lambda map shows clusLerlng of relaLed funcuons. 1he
genome ls 48,314 bp.
27.6 1he Lyuc Cycle uepends on AnuLermlnauon by pn
1he genes concerned wlLh slmllar funcuon are clusLered.
$8 ls an anuLermlnauon
facLor LhaL allows 8nA
polymerase Lo conunue
Lranscrlpuon pasL Lhe
ends of Lhe Lwo
lmmedlaLe early genes.
$9 ls Lhe producL of a
delayed early gene and
ls an anuLermlnaLor LhaL
allows 8nA polymerase
Lo Lranscrlbe Lhe laLe
genes.
llgure 27.09
27.6 1he Lyuc Cycle
uepends on
AnuLermlnauon by pn
27.7 Lysogeny ls MalnLalned by Lhe Lambda 8epressor
roLeln
1he 3&:;7& "($"(..#", encoded by Lhe 02 gene, ls requlred Lo
malnLaln lysogeny.
1he lambda repressor blnds Lo 4
L
and 4
8
#$("&4#". Lo block 8nA
polymerase blndlng Lo promoLers of lmmedlaLe early genes, /
and 0(1 ! block lyuc cycle
1he lmmedlaLe early genes Lrlgger a regulaLory cascade, as a
resulL, Lhelr represslon prevenLs Lhe lyuc cycle from proceedlng.
llgure 27.11
27.7 Lysogeny ls
MalnLalned by Lhe
Lambda 8epressor
roLeln
llgure 27.12
1he lambda
repressor funcuons
as a dlmer.
27.9 1he unA-8lndlng lorm of Lhe Lambda 8epressor
ls a ulmer
A repressor monomer has Lwo dlsuncL domalns.
1he n-Lermlnal domaln conLalns Lhe unA-blndlng slLe.
1he C-Lermlnal domaln dlmerlzes.
8lndlng Lo Lhe operaLor requlres Lhe dlmerlc form so LhaL Lwo
unA-blndlng domalns can conLacL Lhe operaLor
slmulLaneously.
llgure 27.14
lnducuon lf
cleaved
27.10 Lambda 8epressor uses a Pellx-1urn-Pellx Mouf
Lo 8lnd unA
Lach unA-blndlng reglon ln Lhe repressor conLacLs a half-slLe
ln Lhe unA.
1he unA-blndlng slLe of Lhe repressor lncludes Lwo shorL o-
hellcal reglons LhaL L lnLo Lhe successlve Lurns of Lhe "%51(
+(116# of unA (%(3,<=4-"2=%(3,<).
A unA-blndlng slLe ls a (parually) pallndromlc sequence of 17
bp.
llgure 27.16: 1he operaLor ls a 17 bp sequence wlLh an axls of symmeLry Lhrough Lhe
cenLral base palr. Lach half-slLe ls marked by gray arrows.
27.11 Lambda 8epressor ulmers 8lnd Cooperauvely Lo
Lhe CperaLor
8epressor blndlng Lo one operaLor lncreases Lhe amnlLy for
blndlng a second repressor dlmer Lo Lhe ad[acenL operaLor.
1he amnlLy ls 10 greaLer for 4
L
1 and 4
8
1 Lhan oLher
operaLors, so Lhey are bound rsL.
CooperauvlLy allows repressor Lo blnd Lhe 4
L
274
8
2 slLes aL
lower concenLrauons.
llgure 27.22: When Lwo lambda repressor dlmers blnd cooperauvely, each of Lhe subunlLs
of one dlmer conLacLs a subunlL ln Lhe oLher dlmer.
27.11 Lambda 8epressor ulmers 8lnd Cooperauvely Lo
Lhe CperaLor
llgure 27.21: Lach operaLor conLalns Lhree repressor-blndlng slLes and overlaps wlLh Lhe promoLer aL
whlch 8nA polymerase blnds.
27.13 Cooperauve lnLeracuons lncrease Lhe SensluvlLy
of 8egulauon
8epressor dlmers bound aL 4
L
1 and 4
L
2 lnLeracL wlLh dlmers
bound aL 4
8
1 and 4
8
2 Lo form ocLamers.
1hese cooperauve lnLeracuons allows low concenLrauon of
repressor blnds Lo all four operaLor slLes.
When Lhe four operaLor slLes are occupled by repressor, 8nA
pol blnds Lo C
8
3 and acuvaLe Lranscrlpuon of 02 gene.
Cnce repressor ls expressed aL hlgh level, lL blnds Lo C
8
3 and
lnhlblL lLs own Lranscrlpuon: auLoregulaLory!
llgure 27.24
llgure 27.23
27.14 1he 022 and 0222 Cenes Are needed Lo LsLabllsh
Lysogeny
1he delayed early gene producLs cll and clll are necessary for
8nA polymerase Lo lnluaLe Lranscrlpuon aL Lhe promoLer 8
9:
.
cll acLs dlrecLly aL Lhe promoLer and clll proLecLs cll from
degradauon.
1ranscrlpuon from 8
9:
leads Lo synLhesls of repressor and also
blocks Lhe Lranscrlpuon of 0(1.
llgure 27.26
27.13 Lysogeny
8equlres Several
LvenLs
llgure 27.27
1. LsLabllshmenL: cl, cll,
clll, n
2. MalnLenance: cl
27.16 1he Cro 8epressor ls needed for Lyuc lnfecuon
! repressor and Cro can blnd Lo any one of slx operaLors ;!&<
$!=#(#,& %>,!&*.
When Cro blnds Lo 4
8
3, lL prevenLs 8nA polymerase from
blndlng Lo 8
8M
and blocks Lhe malnLenance of repressor
promoLer, l.e., lnhlblLs repressor expresslon (02 gene).
- Amnlues for ! repressor: C
8
1 > C
8
2 = C
8
3
- 8lndlng of repressor Lo C
8
1 and C
8
2 ls cooperauve.
- Amnlues for Cro: C
8
3 > C
8
2 = C
8
1
27. 17. WhaL ueLermlnes Lhe 8alance 8eLween
Lysogeny and Lhe Lyuc Cycle?
1he delayed early sLage when boLh Cro and repressor are belng
expressed ls common Lo lysogeny and Lhe lyuc cycle.
1he crlucal evenL ls wheLher 022 causes sumclenL synLhesls of repressor
Lo overcome Lhe acuon of Cro, cll lnduces ! repressor (cl).

- Upon infection, P
R
expresses
cro and P
L
expresses N.
! Cro expression favors lytic
replication.
- pN induces cII and cIII ! cII
activates P
RE
, resulting in cI
expression.
! cII expression favors
lysogenic growth.
>&?#" "('-3&4#"1 $"#4(,2.
! repressor
- encoded by 0l gene
- 0%, ?1&< %0@6%&# %,$ (#A(#-- &(%,-0(!A@1,. $#-A!&# !&- ,%"#
- represslon by occupauon of slLes overlapplng Lhe promoLer and
excluslon of 8nA polymerase (Lwo lmmedlaLe early genes LhaL
are requlred for lyuc cycle)
- acuvauon by recrulLmenL of 8nA polymerase (lLs own gene)
Cro
- only represses Lranscrlpuon
- ls a slngle-domaln proLeln and blnds as a dlmer Lo unA,
uslng a <#)!BC&D(,C<#)!B "1@E
pn
- AnuLermlnauon facLor (anuLermlnaLor).
- lL allows 8nA polymerase Lo conunue Lranscrlpuon pasL Lhe ends
of Lhe Lwo lmmedlaLe early genes (l.e., 0(1, /).