Plasmids can replicate independently of the host chromosome using their own origin of replication. The two main modes of plasmid replication are theta replication and rolling circle replication. Plasmid replication, host range, copy number, and incompatibility are controlled. The ColE1 plasmid replicates using RNA primer-dependent theta replication controlled by antisense RNA regulation. RNA I binds to RNA II to interfere with its structure and ability to prime replication. The Rop protein enhances binding of RNA I to RNA II to regulate ColE1 copy number.
Plasmids can replicate independently of the host chromosome using their own origin of replication. The two main modes of plasmid replication are theta replication and rolling circle replication. Plasmid replication, host range, copy number, and incompatibility are controlled. The ColE1 plasmid replicates using RNA primer-dependent theta replication controlled by antisense RNA regulation. RNA I binds to RNA II to interfere with its structure and ability to prime replication. The Rop protein enhances binding of RNA I to RNA II to regulate ColE1 copy number.
Plasmids can replicate independently of the host chromosome using their own origin of replication. The two main modes of plasmid replication are theta replication and rolling circle replication. Plasmid replication, host range, copy number, and incompatibility are controlled. The ColE1 plasmid replicates using RNA primer-dependent theta replication controlled by antisense RNA regulation. RNA I binds to RNA II to interfere with its structure and ability to prime replication. The Rop protein enhances binding of RNA I to RNA II to regulate ColE1 copy number.
Plasmids must be able to replicate independently of
the host chromosome. They have their own origin of replication (ori).
There are two general modes of replication:
1) THETA Replication -- which is found in most of the gram negative bacterial plasmids such as ColE1, RK2, and F 2) ROLLINGCIRCLE Replication -- which is found in some gram positive bacterial plasmids such pLS1, pLB4 Replication is central to control number of important plasmid properties:
HOST RANGE, COPY NUMBER, INCOMPATIBILITY and
Plasmid Host Range
Refers to plasmids ability to survive/replicate in a particu
host.
Some plasmids are able to replicate in a limited numbe
bacterial species; they have a NARROW host range.
Examples: ColE1, pBR322, pUC18 plasmids which are lim
to E.coli and some closely related species. Other plasmids are able to replicate in a wide range of bacterial species; they have a BROAD host range. Eg: RSF1010 and RK2
Copy number control of
ColE1 Plasmid
The number of copies of a plasmid can vary from 1 ( F
plasmid) to 100 (pUC18). Copy number is an important property of the plasmid and depends on the mechanism by which it regulates its own replication. Replication control of ColE1 Plasmid ColE1 is a small circular plasmid which codes for a 57 kDal protein toxin (colicin E1) which can kill other E.coli cells.
It carries (oriV) origin of replication and a region
(bom) at which it can be mobilized for transfer to other bacteria.
Other Genes of ColE1 Plasmid and their
functions as follows:
Circular Map of ColE1 Plasmid
Replication of ColE1 proceeds unidirectionally from
oriV and depends on RNA II which is transcribed from a promoter 555 bp away from oriV. When this transcript reaches oriV, one of two possible fates is possible: The RNA-DNA hybrid is cleaved by RNaseH to generate a free 3'-OH end which serves as a primer for DNA polymerase I to initiate replication. Then DNA polymerase III takes over after a short distance.
If RNA-DNA hybrid is NOT cleaved by RNaseH and
transcription continues nonproductively. These fates depends on the formation of a specific secondary structure in the nascent RNA II transcript; If the structure forms correctly then RNaseH processing occurs correctly. If anything interfere with the formation of correct secondary structure will interfere with replication
A specific RNA called RNA I whose function is to
interfere with the secondary structure of RNA II
This RNA I is a 108 nt molecule which is transcribed in the
opposite direction to RNA II from a promoter located between the RNA II promoter and the origin of replication
Since RNA I is complementary to RNA II, it form a
stable RNA-RNA hybrid with it. If it does, then RNA II will not be able to function as a primer.
This is an example of antisense RNA
regulation.
Replication of ColE1 also depends on the Rop
protein
Rop is a small protein (63 aas) which increases the
rate of binding of RNA I to RNA II. If Rop is absent, plasmid replication will be more frequent.