分子生物學(xué) 第十一章 噬菌體的戰(zhàn)略（Phage strategies）

1、Chapter 11,Phage strategies,11.1 Introduction11.2 Lytic development is divided into two periods11.3 Lytic development is controlled by a cascade11.4 Functional clustering in phages T7 and T411.5 Lambda immediate early and delayed genes are needed for both lysogeny and the lytic cycle11.6 The lytic c

2、ycle depends on antitermination11.7 Lysogeny is maintained by repressor protein11.8 Repressor maintains an autogenous circuit11.9 The repressor and its operators define the immunity region11.10 The DNA-binding form of repressor is a dimer 11.11 Repressor uses a helix-turn-helix motif to bind DNA11.1

3、2 Repressor dimers bind cooperatively to the operator11.13 Repressor at OR2 interacts with RNA polymerase at PRM11.14 The cII and cIII genes are needed to establish lysogeny11.15 PRE is a poor promoter that requires cII protein11.16 Lysogeny requires several events11.17 The cro repressor is needed f

4、or lytic infection11.18 What determines the balance between lysogenic and the lytic cycle?,Episome is a plasmid able to integrate into bacterial DNA. EpistasisImmunity in phages refers to the ability of a prophage to prevent another phage of the same type from infecting a cell. It results from the s

5、ynthesis of phage repressor by the prophage genome.Induction refers to the ability of bacteria (or yeast) to synthesize certain enzymes only when their substrates are present; applied to gene expression, refers to switching on transcription as a result of interaction of the inducer with the regulato

6、r protein.Lysogeny describes the ability of a phage to survive in a bacterium as a stable prophage component of the bacterial genome.Lytic infection of bacteria by a phage ends in destruction of bacteria and release of progeny phage.Plasmid is an autonomous self-replicating extrachromosomal circular

7、 DNA.Prophage is a phage genome covalently integrated as a linear part of the bacterial chromosome.,11.1 Introduction,Figure 11.1 Lytic development involves the reproduction of phage particles with destruction of the host bacterium, but lysogenic existence allows the phage genome to be carried as pa

8、rt of the bacterial genetic information.,11.1 Introduction,Figure 11.2 Several types of independent genetic units exist in bacteria.,11.1 Introduction,Figure 11.3 Lytic development takes place by producing phage genomes and protein particles that are assembled into progeny phages.,11.2 Lytic develop

9、ment is controlled by a cascade,Figure 11.4 Phage lytic development proceeds by a regulatory cascade, in which a gene product at each stage is needed for expression of the genes at the next stage.,11.2 Lytic development is controlled by a cascade,Figure 9.31 Switches in transcriptional specificity c

10、an be controlled at initiation or termination.,11.2 Lytic development is controlled by a cascade,Figure 11.5 Phage T7 contains three classes of genes that are expressed sequentially. The genome is 38 kb.,11.3 Functional clustering in phages T7 and T4,Essential genes are indicated by numbers. Nonesse

11、ntial genes are identified by letters. Only some representative T4 genes are shown on the map.,11.3 Functional clustering in phages T7 and T4,Figure 11.6 The map of T4 is circular. There is extensive clustering of genes coding for components of the phage and processes such as DNA replication, but th

12、ere is also dispersion of genes coding for a variety of enzymatic and other functions.,Figure 11.7 The phage T4 lytic cascade falls into two parts: early and quasi-late functions are concerned with DNA synthesis and gene expression; late functions are concerned with particle assembly.,11.3 Functiona

13、l clustering in phages T7 and T4,Figure 11.24 RNA polymerase binds to PRE only in the presence of CII, which contacts the region around -35.,11.3 Functional clustering in phages T7 and T4,Figure 11.8 The lambda lytic cascade is interlocked with the circuitry for lysogeny.,11.4 The lambda lytic casca

14、de relies on antitermination,Figure 11.9 The lambda map shows clustering of related functions. The genome is 48,514 bp.,11.4 The lambda lytic cascade relies on antitermination,Figure 11.10 Phage lambda has two early transcription units; in the leftward unit, the upper strand is transcribed toward th

15、e left; in the rightward unit, the lower strand is transcribed toward the right. Promoters are indicated by the shaded red or blue arrowheads. Terminators are indicated by the shaded green boxes. Genes N and cro are the immediate early functions, and are separated from the delayed early genes by the

16、 terminators. Synthesis of N protein allows RNA polymerase to pass the terminators tL1 to the left and tR1 to the right.,11.4 The lambda lytic cascade relies on antitermination,Figure 11.11 Lambda DNA circularizes during infection, so that the late gene cluster is intact in one transcription unit.,1

17、1.4 The lambda lytic cascade relies on antitermination,Immunity in phages refers to the ability of a prophage to prevent another phage of the same type from infecting a cell. It results from the synthesis of phage repressor by the prophage genome.Virulent phage mutants are unable to establish lysoge

18、ny.,11.5 Lysogeny is maintained by an autogenous circuit,Figure 11.12 The lambda regulatory region contains a cluster of trans-acting functions and cis-acting elements.,11.5 Lysogeny is maintained by an autogenous circuit,Figure 11.13 Wild-type and virulent lambda mutants can be distinguished by the

19、ir plaque types. Photograph kindly provided by Dale Kaiser.,11.5 Lysogeny is maintained by an autogenous circuit,Figure 11.14 Lysogeny is maintained by an autogenous circuit (upper). If this circuit is interrupted, the lytic cycle starts (lower).,11.5 Lysogeny is maintained by an autogenous circuit,

20、Figure 11.14 Lysogeny is maintained by an autogenous circuit (upper). If this circuit is interrupted, the lytic cycle starts (lower).,11.5 Lysogeny is maintained by an autogenous circuit,Figure 11.15 The N-terminal and C-terminal regions of repressor form separate domains. The C-terminal domains ass

21、ociate to form dimers; the N-terminal domains bind DNA.,11.6 The DNA-binding form of repressor is a dimer,Figure 11.16 Repressor dimers bind to the operator. The affinity of the N-terminal domains for DNA is controlled by the dimerization of the C-terminal domains.,11.6 The DNA-binding form of repre

22、ssor is a dimer,Figure 11.16-2 Lambda repressor binds to operators with second-order kinetics.,11.6 The DNA-binding form of repressor is a dimer,Figure 11.17 Lambda repressors N-terminal domain contains five stretches of a-helix; helices 2 and 3 are involved in binding DNA.,11.7 Repressor binds coop

23、eratively at each operator using a helix-turn-helix motif,Figure 11.18 In the two-helix model for DNA binding, helix-3 of each monomer lies in the wide groove on the same face of DNA, and helix-2 lies across the groove.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix mot

24、if,Figure 11.19 Two proteins that use the two-helix arrangement to contact DNA recognize lambda operators with affinities determined by the amino acid sequence of helix-3.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix motif,Figure 11.20 A view from the back shows that

25、the bulk of the repressor contacts one face of DNA, but its N-terminal arms reach around to the other face.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix motif,Figure 11.21 Each operator contains three repressor-binding sites, and overlaps with the promoter at which RN

26、A polymerase binds. The orientation of OL has been reversed from usual to facilitate comparison with OR.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix motif,Figure 11.21 Each operator contains three repressor-binding sites, and overlaps with the promoter at which RNA p

27、olymerase binds. The orientation of OL has been reversed from usual to facilitate comparison with OR.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix motif,Figure 11.14 Lysogeny is maintained by an autogenous circuit (upper). If this circuit is interrupted, the lytic cyc

28、le starts (lower).,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix motif,Figure 11.22 Positive control mutations identify a small region at helix-2 that interacts directly with RNA polymerase.,11.7 Repressor binds cooperatively at each operator using a helix-turn-helix m

29、otif,Figure 11.12 The lambda regulatory region contains a cluster of trans-acting functions and cis-acting elements.,11.8 How is repressor synthesis established?,Figure 11.23 Repressor synthesis is established by the action of CII and RNA polymerase at PRE to initiate transcription that extends from

30、 the antisense strand of cro through the cI gene.,11.8 How is repressor synthesis established?,Figure 11.24 RNA polymerase binds to PRE only in the presence of CII, which contacts the region around -35.,11.8 How is repressor synthesis established?,Figure 11.25 Positive regulation can influence RNA p

31、olymerase at either stage of initiating transcription.,11.8 How is repressor synthesis established?,Figure 11.26 A cascade is needed to establish lysogeny, but then this circuit is switched off and replaced by the autogenous repressor-maintenance circuit.,11.8 How is repressor synthesis established?

32、,Figure 11.19 Two proteins that use the two-helix arrangement to contact DNA recognize lambda operators with affinities determined by the amino acid sequence of helix-3.,11.9 A second repressor is needed for lytic infection,Figure 11.27 The lytic cascade requires Cro protein, which directly prevents repressor maintenance via PRM, as well as turning off delayed early gene expression, indirectly preventing repressor establishment,11.9 A second repressor is needed for lytic infection,Figure