Microbiology 460

Department of Microbiology
Southern Illinois University at Carbondale


Lecture Topic: Integrons and Mobile Gene Cassettes


Reference: Hall and Collis. 1995. Molec. Microbiol. 15:593-600.


Integron - genetic unit that includes genes of a site-specific recombination system capable of capturing and mobilizing genes contained in mobile elements called gene cassettes.
Integron structure consists of two conserved regions (intI regions and sulI region) flanking a variable region (gene cassette) which contains one or more resistance genes. Cassette-free integrons exist in which the int and sul regions are contiguous.
The intI region encodes integrase.
The sulI region confers resistance to sulphonamides.
The gene cassette is mobile and can also exist in free circular form. However, these cassettes do not include all functions required for their mobility. Cassettes are only formally part of the integron when they are integrated at the integron-receptor site. The genes within the cassette are promoterless.
It is interesting to note that this is thus a site-specific recombination system in which the integrase function is supplied by the recipient (the integron) and not the mobile insertion unit (the gene cassette).
The inserted genes of the mobile cassette are all in the same orientation with respect to the int (5'-CS [conserved sequence]) and sul (3'-CS) regions. This orientation allows them to be transcribed from a common promoter, Pant, that is located in the 5'-CS and is responsible for transcription of the antibiotic-resistance genes.
When more than one cassette is present, the cassette-encoded genes are co-transcribed from Pant. However, downstream genes are transcribed less frequently due to premature termination of transcripts.
The recombination site that acts as the receptor site for insertion of the cassettes is a 40 to 70-base element called attI. This site spans the 5'-CS and 3'-CS junction and the distal portions lie within these conserved regions. The site within the cassette that interacts with the attI site is called a 59-base element which in a circular free cassette consists of two inverted repeats.
Loss of cassettes is dependent on integrase function and excision results in circular free cassettes. These free cassettes are not able to replicate but are able to participate in integration into another integron, thus spreading the antibiotic resistance genes from one integron to another.
The insertion/excision events mediated by the integron-encoded integrase can lead to cointegrate formation between two integron-containing plasmids present in the same cell and the subsequent resolution of these cointegrates. If cointegrate resolution occurs at sites other than those used for integration, a reciprocal acquisition and loss of gene cassettes can occur resulting in one cassette-free integron and one integron with two integrated cassettes. This may be significant if the new recipient host plasmid is transferrable or has a broader host range.
No information is as yet available on the origin of gene cassettes. One hypothesis is that they arose from reverse transcription of a mRNA, which would explain why gene cassettes contain genes with no flanking sequences such as promoters. The 59-base element may have originated from a transcription terminator (a hairpin loop) and in fact recent evidence indicates that these elements can indeed function as terminator sites.


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Last updated: 28-Oct-02 / laa