gene expression and replication in medium dna viruses
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What can the medium sized viruses do that the small ones can not What can the medium sized viruses do that the small ones can not? Adenovirus: more sophisticated gene expression strategies Lambda: two pathways for infectionTRANSCRIPT
Gene Expression and Replication in Medium DNA Viruses
Phage Lambda Adenovirus What can the medium sized viruses do that
the small ones can not
What can the medium sized viruses do that the small ones can
not?Adenovirus: more sophisticated gene expression strategies
Lambda: two pathways for infection Phage Lambda Bacteriophage
isolated from human gut-Andre Lvov?
dsDNA, 48.5 kbp Linear, sticky ends cos Complex capsid Unusual
biological properties noticed Model system from 1950s Cloning
vector Evergreen State-Olympia Mansfield State (Ohio)
The Evergreen State College Phage Lab has been a center for
undergraduate research at Evergreen since Betty Kutter came here in
1972, one year after the college opened. Today, there are generally
students involved in work in the lab under the direction of Kutter
and faculty colleague Andrew Brabban. We are focusing particularly
on phage ecology, with the aims of understanding the infection
process under anaerobic and other conditions better reflecting
those in the natural environment, as well as on interactions
between phages of different families during simultaneous infection
and the building of cocktails for potential therapeutic
applications. Dr. Elizabeth Kutter: Dr. Andrew Brabban: Evergreen
State-Olympia Mansfield State (Ohio) Bacteriophage Therapy
Inst-Tbilisi Bacteriophage therapy was used broadly in the Soviet
Union, particularly the Republic of Georgia, which has been the
global center of phage expertise for over 80 years. Lambda Entry
and Uncoating
Non-contractile Lambda J proteinuses Lam B as receptor DNA enters
through mannose permease complex Lambda genetic map Shortly after
entry the linear DNA forms a circle
Cos sites are complementary with one another Assembles functional
regions Lambdas two developmental pathways
In some cases lambda follows the usual pathway in which the host
cell is killed straightaway-the lytic pathway. In other cases
lambda DNA inserts into the bacterial chromosome to form a
prophage. The prophage remains in the chromosome as a passenger
until the host cell is stressed or induced. Then it lyses the host
cell to release new phage. The latter pathway is called the
lysogenic or prophage pathway. (temperate, vegetative) Lysis and
Lysogeny for Phage Lambda
What are the advantages of lysogeny for lambda? How does lambda
make its decision for lysis or lysogeny? Phage lambda regulatory
region
Key region regulates developmental decision P = promoter O =
operator T = terminator Operon Model Shown for lactose operon
Repressor protein binds to operator sequence: No mRNA so gene is
off! Inducer causes shape change in repressor Repressor drops off
operator mRNA produced so gene is on Gene Expression Immediately
after infection, N and cro are transcribed by host RdRp (immediate
early genes) N is an antiterminator- it allows transcription and
expression of cIII, cII, O, Q (early genes). Cro is a repressor. It
can block transcription from Pr and Pl. N says go, cro says stop
Lytic pathway If cro builds up quickly enough it prevents further
synthesis of N and of cro itself. Q allows expression of late genes
that encode for lytic functions. Establishment of Lysogeny
If cII builds up fast enough it activates the transcription of cI
and also some integration functions. cI is called the lambda
repressor. It shuts off all lambda genes except itself and allows
lambda DNA to integrate into the host chromosome. Lysis and
Lysogeny Compete
If cro wins then late genes are expressed and lysis follows. If N,
cII, cIII win then the lambda repressor is expressed from cI and
lysogeny follows. This medium-sized virus is complex enough for two
developmental options. Post Script:Lambda repressor binding
Binds to operator as a homodimer Helix-turn-helix protein Up to 3
dimers can bind Binding is cooperative Cooperative repressor
binding produces
steep response curve-all or nothing response R E P R E S S I O N
REPRESSOR CONCENTRATION When enough repressor is around is turns
off its own synthesis
When repressor levels drop-more is made Induction involves
destruction of repressor Superinfection immunity Lambda DNA
Replication
Lambda uses a combination of host and viral proteins. Theta
strategy for first few rounds of replication. Then shifts to
rolling circle. Long tail converted to ds concatemer. Cleavage
occurs at cos sites during packaging.