who cares about rho gtpases? bordetella spp.neisseria spp. clostridium spp. salmonella spp

Who cares about Rho GTPases?

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Bordetella spp. Neisseria spp.

Clostridium spp.

Salmonella spp.

Historical GTPase Events

• 1985 – isolation of Rho = ‘Ras homolog’

• 1989 – C3 transferase shown to inactivate RhoC => actin disruption in host cells. Isolation of Rac = ‘Ras substrate of C3’.

• 1990’s – Rho/Rac GTPases shown to act as switches to control membrane receptors and actin cytoskeleton plasticity.

Salmonella enterica

Type III Secretion System (TTSS) for translocation of virulence factors

http://info.med.yale.edu/micropath/galan/Pages/galan_overview.html

Salmonella chaperone proteins

…after effector translocation…Bacterial changes: transient bacterial ‘invasome’ appendages

Host cell changes: macropinocytosis for uptake of Salmonella in nonphagocytic cells

bacterial induced apoptosis in phagocytic cells.

Salmonella invasion summary

http://info.med.yale.edu/micropath/galan/Pages/galan_overview.html

Bacterial StrategiesRHO FAMILY GTPASES: Rho, Rac, Cdc42

BACTERIAL INHIBITION OF RHO PROTEINS:

-Large clostridial toxins: Toxin A, B, Lethal Toxin

-C. botulinum C3 transferase

-Salmonella SptP, Yersinia YopE

BACTERIAL ACTIVATION OF RHO PROTEINS:

-E.coli CNF1 and 2 Toxins

-Bordetella dermonecrotizing toxin (DNT)

-Salmonella SopE, Yersinia CNFY

Fig. 1. Rho GTPases are targets for bacterial virulence factors.

Fig. 2. Bacterial virulence factors affect spatial and temporal regulation of Rho.

Why target Rho GTPases?

• Invasion can be dangerous!– Innate immunity – recognize ‘non-self’ and

opsonize for phagocytic cell recognition. LPS recognized by TLRs stimulates NF-KB and leads to transcription of antibacterial factors.

– Cell shedding removes adhered bacteria.– Adaptive immunity… takes time.

• Virulence factors help microbes invade on their own terms! Rho GTPases are key.

Fig. 3. Virulence factors can adapt or mimic eukaryotic mechanisms.

How do virulence factors enter?

• Toxins– Can act distantly to bacteria because all required

elements for virulence self-contained.– Diptheria A-B example:

• A region – catalytically active, delivered to cytosol.• B region – for binding host cell and translocating the

A-enzymatic fragment to host cytosol at low pH.

• Type III or IV Secretion Systems

Fig. 4. Domain organization of virulence factors activating or inhibiting Rho GTPases.

Fig. 5. Mechanisms to transfer Rho virulence factors into the cell cytosol.

Rho Inhibitors

• Classical model:– Toxins effects were irreversible, while TTSS

induced reversible changes in Rho.

• Large clostridial toxins (LCTs)– Toxins A, B, Lethal Toxin.

• Type Three Secretion Systems– Pseudomonas ExoS, ExoT.– Salmonella SptP, Yersinia YopT, YopE.

• C3 transferases, YopT: spatial regulation.

Rho activators• E.coli CNF, Bordetella DNF:

– Block RhoGAP activity so GTPase is permanently active until ubiquitinylation and proteosomal degradation.

– Is proteosomal degradation of overactivated Rho a cellular defense that microbes are taking advantage of?

• Salmonella SopE, E2:– Rho GEF function to activate Rho but is

counterbalanced by SptP GAP activity.

Fig. 5. Comparison of activation-deactivation of Rac by Salmonella SopE/Sptp and E. coli CNF1.

Why activation/deactivation?

• Whether the bacteria supplies the Rho counterbalance (Salmonella) to virulence or the host cell provides it (E.coli)…

-is it simply to return to ‘normal’ cell function?

-or to enhance bacterial uptake?

-or to avoid non-physiologic cell environs that prevent bacterial uptake at all?

Summary• Rho GTPases can be influenced by:

– Activities from separate bacterial factors• Salmonella SopE, SptP; Yersinia YopE,YopT

– Dual activity factors• Pseudomonas ExoS, ExoT

– Single activity proteins• E. coli CNF1 Toxin

• What is the future of the host-pathogen interaction? Extremes vs balance?

The End