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Printable Handouts
Navigable Slide Index
- Introduction
- Linear DNA in diverse bacteria
- The 'end replication' problem
- Solving the end replication problem
- Streptomyces
- Replication pathway for Adenovirus and Bacillus phage f29
- Streptomyces DNA replication
- A minimal pSLA2 replicon
- Cis and trans acting regions for pSLA2 replication
- Linear DNA replication intermediate
- Predicted secondary structure of S. lividans SLP2 plasmid end
- Patching by terminal protein
- The terminal proteins of Streptomyces
- Comparison of seven TP sequences
- Recruitment of TP
- Deoxynucleotidylation of TP and DNA synthesis
- Tap directs Tpg to its location and adds dCMP
- Streptomyces summary
- Borrelia
- The segmented genome of Borrelia burgdorferi
- Replication pathway for linear plasmids in Borrelia (1)
- Origin mapping by nascent strand analysis
- Origin location in the linear chromosome
- Plasmid maintenance proteins in B. baveriensis
- Minimal replicons
- Regions required for Ip17 replication
- The BBD14 (Family 62) and BBD21 (Family 32) proteins
- The telomere resolvase (ResT)
- Replication pathway for linear plasmids in Borrelia (2)
- Purification of recombinant B. burgdorferi ResT
- Assay for telomere resolution by ResT
- The telomeres
- Three classes of telomere in Bb. Cleavage between bp 3 and 4 in all
- Minimal substrate requirements
- How does DNA cleavage and ligation occur?
- Comparison of domains in 3 telomere resolvases
- Active site residues of telomere resolvases
- Mechanism of action of ResT: a 2-step transesterification
- Features of telomere resolvases
- How does hairpin formation occur?
- Alignment of hairpin binding molecules of Tn5 and Tn10 with ResT
- A composite active site promotes telomere resolution
- A composite active site
- Structure and function of telomere resolvases
- Genome plasticity in Borrelia
- Sequence scrambling in linear plasmids
- Telomere exchanges
- What causes high incidence of gene duplications and telomere exchanges in Borrelia species?
- ResT reversal using a plasmid substrate
- Telomere exchange by ResT-mediated telomere fusions
- Sequence scrambling near the B. burgdorferi telomeres
- Reversal of telomere resolution (telomere fusion)
- Conclusions: genome plasticity and ResT (1)
- Conclusions: genome plasticity and ResT (2)
- The end
Topics Covered
- Linear DNA replication in bacteria
- The ‘end replication’ problem
- Linear DNA replication in Streptomyces
- Linear DNA replication in Borrelia
- Telomere resolvase (ResT)
- Genome plasticity in Borrelia
- Sequence scrambling in linear plasmids
- Reversal of telomere resolution
- Covalently closed hairpin ends
Talk Citation
Chaconas, G. (2024, January 31). Replication of linear plasmids in bacteria [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved November 21, 2024, from https://doi.org/10.69645/YSNK2670.Export Citation (RIS)
Publication History
Financial Disclosures
- There are no commercial/financial matters to disclose.
A selection of talks on Microbiology
Transcript
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0:00
The title of this
presentation is
Replication of Linear
Plasmids in Bacteria.
0:08
Let's begin our
discussion with a look at
the distribution of linear
DNA in the bacterial world.
This figure shows a tree of
relatedness for bacterial phyla,
in which phyla have
linear replicons.
Two things are immediately
obvious from this tree.
First, a few bacterial
phyla carry linear DNA.
Second, the ones that do
are not closely related.
Linear DNA has been found in
protobacteria, actinobacteria,
and spirochetes.
Three phyla that are
evolutionarily distant
from each other.
The observed linear DNA
has been found in the form
of bacterial chromosomes,
plasmids and Phages.
The reasons why linear
DNA in bacteria is
not a common occurrence is
discussed in the next slide.
1:01
Most bacterial plasmids
are circular molecules.
This simplifies the DNA
replication process.
With linear DNA molecules,
we encounter what
has been referred
to as the end
replication problem.
This was first noted by James
Watson in the early 1970s.
The problem results
from the fact
all DNA polymerases require
a primer to initiate
DNA synthesis.
As shown in this illustration,
when synthesis is complete
on the lagging strand,
and the RNA primer at
the five prime end of
the newly replicated
strand is removed,
we are left with a gap.
This gap cannot be filled in
de novo by a DNA polymerase,
and we are left with
unreplicated DNA at
the ends of the lagging strand
of linear DNA molecules.