1. The Evolutionary Sequence of the Anthrax Toxins
Carolyn Vanek
The Evolutionary Sequence of the Anthrax Toxins

2. Anthrax: Background Routes of Infection: Skin Lung Intestines
Spores can stay in the ground for decades
Herbivores eat something containing the spores, they become infected, infection is passed on
2000 cases per year, 2 in the US
Respiratory anthrax has 50-80% mortality rate with treatment

3. The Three Horsemen of the Apocolypse
Protective Antigen
Edema Factor
Lethal Factor
Death
Edema
Death
LF not toxic by itself
Nothing

4. Protective Antigen
Needed for internalization

5. Edema Factor Calmodulin-sensitive adenylate cyclase
Provokes dramatic elevation of intracellular cAMP levels in the host
Needs calmodulin to be active

6. Lethal Factor
t is a protease that cleaves the N-terminal of most dual specificity mitogen-activated protein kinase kinases (MAPKKs or MAP2Ks)

7. Question: What can phylogenetic analysis show about Anthrax’s inheritance of these three proteins?

8. Sequence Obtained from NCBI BLOSUM50 Matrix
Targets alignments with 20-30% Identity
Depending on sequence, retrieve sequences
Removed repeated and hypothetical proteins
Manual curation
Final alignments had anywhere from proteins
Matrix because not that many poisons (thank god) so looking for larger evolutionary differences
Looked at Orthologs primarily
Max Expect value of 1
Frequently found one of the EF in PA. Early indication of duplications

9. Side Note: Lethal Factor

10. Sequence Alignment Used Tcoffee, default Tcoffee best but slowest
Few sequences in analysis

11. Tree Creation Divergent sequences were frequently an issue
Used Observed over Poisson
Tested Including Gaps and Ignoring Gaps
Including Gaps gave better bootstrap on trees
Trees also made more sense in the way proteins were grouped