Contact-dependent growth inhibition

The Type IV Secretion System (T4SS) is found in many species of Gram-negative and Gram-positive bacteria as well as in archea and are typically associated with conjugation or delivery of virulence proteins to eukaryotic cells.[4] Some species of plant pathogen Xanthomonas, however, possess a particular T4SS capable of mediating CDI by delivering a peptidoglycan hydrolase. This effector kills targets that do not have the cognate immunity protein similar to other CDI systems.[5]

The first CDI system to be discovered was a Type V secretion system, encoded by the cdiBAI gene cluster found widespread throughout pathogenic Gram-negative bacteria. The first protein encoded in the operon, CdiB, is an outer membrane beta-barrel protein that exports CdiA, presenting it on the cell surface of a CDI-expressing (CDI+) bacterium. CdiA is predicted to form a filament several nanometers long that extends outward from the CDI+ cell in order to interact with neighbouring bacteria via outer membrane protein receptors to which it will bind.[2] The C-terminal 200-300 amino acids of CdiA harbours a highly variable toxic domain (CdiA-CT), which is delivered into a neighbouring bacterium upon receptor recognition, enabling the CDI+ cell to arrest the growth of the cell into which it delivers this CdiA-CT toxin. This toxic domain is linked to the rest of CdiA via a VENN peptide motif and vary significantly more between species than does the rest of CdiA.[6] CdiI is an immunity protein to prevent auto-inhibition by the C-terminal toxin. This also prevents the bacteria from killing or inhibiting the growth of their siblings as long as these possess the immunity gene.[7] Many CDI systems contain additional cdiA-CT/cdiI pairs called "orphans" following the first copy [8] and these orphans can be connected to different main CdiA:s in a modular fashion.[6]

The Type VI Secretion System (T6SS) is widely spread amongst Gram-negative bacteria and consists of a protein complex, encoded by several different genes, forming "needle-like" structure capable of injecting effector molecules into neighbouring target cells similar to the contractile tail of the T4 bacteriophage. One T6SS may have several different effectors such as PAAR-domain toxins or Hcp toxins and some species can deliver these toxins into both prokaryotes and eukaryotes.[3][9]

The Rearrangement hotspot system (Rhs) exists in both Gram-negative and Gram-positive bacteria. Similar to CdiA, these systems consists of big proteins with a conserved N-terminal domain and a variable C-terminal toxin domain requiring a cognate immunity protein. Many Rhs systems contain PAAR-domains (Proline-Alanine-Alanine-Arginine) which can interact with the VgrG of the T6SS apparatus making it required for Rhs secretion.[3][10] The name Rearrangement hotspots comes from the discovery when the system was first identified as elements on the E. coli chromosome that were continuously rearranging.[11][12] The Gram-positive soil bacterium Bacillus subtilis possesses an Rhs homolog called Wall-associated protein A (WapA) capable of mediating CDI whilst requiring a cognate immunity protein, WapI, to prevent auto-inhibition.[10]

In E. coli, CdiA molecules may interact with those found on neighboring cells, independent of the receptor to which CdiA binds. In addition with receptor binding, these homotypic interactions cause cell-cell aggregation and promote biofilm formation for CDI+ bactera. In a similar fashion, the CdiA homolog BcpA in Burkholderia thailandensis causes up-regulation of genes encoding pili and polysaccharides when delivered to sibling cells which are in possession of the immunity protein BcpI. This change in gene expression leads to increased biofilm formation in the bacterial population through a phenomenon now known as Contact-Dependent Signalling. Furthermore, the T6SS in V. cholerae is active in biofilms, enabling a cell expressing T6SS to kill nearby cells which do not have the specific immunity.[5]