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BACTERIAL

CONJUGATION
INTRODUCTION
Bacteria posses basic three mechanisms of lateral or horizontal gene
transfer. They are,
 Transformation- transfer of gene from one bacterium to other
without cell to cell contact.
 Transduction– transfer of gene from one bacterium to other via
agency of phages.
 Conjugation- transfer of gene from one bacterium to other by cell
to cell contact by conjugal pore.
Conjugation was first discovered in 1946 by two renowned
scientists of the time Joshua Lederberg and Edward Tatum.
Bacterial conjugation is often incorrectly regarded as the
bacterial equivalent of sexual reproduction or mating. Bacterial
conjugation is merely a transfer of gene from one bacterium to
other unlike the fusion of gametes in sexual reproduction in higher
organisms.
DISTINGUISHING CHARACTERISTICS OF
CONJUGATION

 DNA transfer requires cell-cell contact.


 DNA transfer occurs via a conjugal pore.
 DNA transfer occurs in one direction - from donor to
recipient not vice versa
 DNA transfer does not require protein synthesis in donor.
 DNA transfer requires energy in donor cell - primarily ATP.
The F+ strain

The mating bridge

The F- strain
Fertility plasmid
The E. coli fertility plasmid are most extensively studied plasmid.
The general features of this plasmid are,
1. Large circular plasmid (100 kb)
2. Only 60% genes has been mapped.
3. 32 kb is organized as a unit to transfer its genome to another
bacteria (transfer region or tra genes)
4. Two methods of replication:
a. oriV as free plasmid (one copy/ bacterial chromosome)
b. uses bacterial chromosomal origin when integrated (oriC); oriV is
suppressed.
mechanism
 Plasmid replication requires a "mating bridge" between the donor
and recipient cells.
Before the mating bridge can form:
- donor must recognize recipient cell
- donor must make contact with recipient cell
The conjugative functions of the F plasmid are specified by a
cluster of at least 25 transfer (tra) genes. They determine:
 expression of F pili.
 synthesis and transfer of DNA during mating
 interference with the ability of F+ bacteria to serve as recipients.
Mechanism contd…
Each F+bacterium contains F pili

Binding of F pili to specific outer membrane protein

Intercellular cytoplasmic bridge formation

Transfer of single strand from donor to recipient


Transferred strand is
The copy is retained in donor converted into double
stranded circle in recipient.
The newly synthesized double stranded
DNA is called exogenote and native DNA
as endogenote.
Domain interacting with outer
membrane protein

3D X ray crystallographic structure of protein pilin


General representation of mechanism of conjugation
The relaxase and the relaxosome
 When conjugation is initiated, via a mating signal, a relaxase
enzyme (an endonuclease) creates a nick in one plasmid DNA
strand at the origin of transfer, or oriT.
 The relaxase may work alone or in a complex of over a dozen
proteins, known collectively as a relaxosome.
 In the F-plasmid system, the relaxase enzyme is called TraI and
the relaxosome consists of TraI, TraD, TraY, TraM, and the
integrated host factor, IHF.
 The transferred, or T-strand, is unwound from the duplex plasmid
and transferred into the recipient bacterium in a 5'-terminus to
3'-terminus direction.
 The remaining strand is replicated, either independent of
conjugative action (vegetative replication, beginning at the oriV)
or in concert with conjugation (conjugative replication similar to
the rolling circle replication of lambda phage).
Mechanism illustrated

Initiates
F plasmid
transfer

Helps creating
the ssDNA
conformation

Sterically
prepare the DNA
for other
proteins to
interact in
specific spots.
Conjugation protein assembly
Domains of protein trai
Formation of Hfr strain

There are two mechanisms of


integration
-homologous recombination
-transposition
Formation of F’ plasmid

1. Plasmids that “leave” the genome carrying chromosomal DNA are known
as prime factors.
2. They “leave” the chromosome by homologous recombination,
resulting in a deletion in the chromosome
Animations
Host range
 Bacteria (same or different species)
 Yeast
 Plant cells
 Mammalian cells
conclusion

The genetic information transferred is often beneficial to the


recipient cell. Benefits may include antibiotic resistance, other
xenobiotic tolerance, or the ability to utilize a new metabolite.
Such beneficial plasmids may be considered bacterial
endosymbionts. Some conjugative elements may also be viewed
as genetic parasites on the bacterium, and conjugation as a
mechanism was evolved by the mobile element to spread itself
into new hosts.