1 Emily Pedersen

Micrb 311
Dr. Marina Lazic

A Secret Double Agent Lying Within S. aureus Virulence Systems

Staphylococcus aureus. The most dangerous species of pathogen. I’m kidding; it's not even close to the

most dangerous. It is, however, a gram positivegram-positive pathogen that I wasn’t allowed to deal with

in my Microbiology 265 lab for fear of getting sick. This pathogen is quite well known, existing in both

soil and the human skin. S. aureus is one of the few bacteria that has a complete tricarboxylic acid (TCA)

cycle1. An article published in 2003 studies a defect present in some strains of S. aureus that affects the

functioning of the TCA cycle in regards toregarding growth and virulence2, with the goal of determining

whether there are physiological consequences to said mutation.

First, some background so we can understand the

role the impairment plays in virulence. Within S.

aureus cells, a complete TCA cycle is utilized to

catabolize acetate, leading to the creation of

multiple virulence factors in the post-exponential

phase of growth. Aconitase, an important enzyme

in the TCA cycle and thus a pre/postcursor of acetate catabolism, is directly correlated with an increased

secretion of virulence factors. These virulence factors, furthermorefurthermore, referred to as S. aureus’s

weapons of destruction, include α-toxin, β-hemolytic activity, and protein A. In post exponential phase,

the carbon and glucose supply has run dry, leading gram positivegram-positive pathogens to switch gears

and employ the TCA cycle to generate citrate 2. Acetate is subsequently catabolized by aconitase, allowing

the expression of genes related to the weapons of destruction. If aconitase acitivtyactivity is impaired,

there will be a severe decrease in virulence factor expression, and thus virulence itself.
1:Kwong WK, Zheng H, Moran NA. 2017. Convergent evolution of a modified , acetate-driven TCA cycle in bacteria. Nat Microbiol. 2:17067.
DOI: 10.1038/nmicrobiol.2017.67
2:Somerville GA, Said-Salim B, Wickman JM, Raffel SJ, Kreiswirth BN, Musser JM. 2003. Correlation of Acetate Catabolism and Growth Yield
in Staphylococcus aureus: Implications for Host-Pathogen Interactions. Infect and Immun. 71(8): 4724-4732. DOI: 10.1128/iai.71.8.4724-
4732.2003
Image 1:Somerville GA, Said-Salim B, Wickman JM, Raffel SJ, Kreiswirth BN, Musser JM. 2003. Correlation of Acetate Catabolism and
Growth Yield in Staphylococcus aureus: Implications for Host-Pathogen Interactions. Infect and Immun. 71(8): 4724-4732. DOI:
10.1128/iai.71.8.4724-4732.2003
2

Alright now to the main topic - the double agent(s). Within the NCTC 8325 and N315 strains of S.

aureus, there is a defect that causes decreased aconitase activity, effectively reducing the pathogens

growth rate and virulence. This has led to the hypothesis that secondary metabolite catabolism can be lost.

The decreased aconitase activity is caused by an impairment that has not been pinpointed, though there

are many genes I’ll discuss as possible traitors. To deal with this loss of function, the two strains should

compensate by increasing the catabolism of other metabolites, right? Wrong. The two strains have not

shown a significant difference in amount of ammonia, a possible replacement, made. This means that the

double agents have affectively diminished the growth rate of strains NCTC 8325 and N315. But have the

double agents affected the weapons of destruction as well? The results show that they didn’t. While itsit’s

reasonable to assume that the double agents would not only affect the spread of the pathogen, but also the

destructiveness of it, virulence factors in the two strains did not show significant differences in

presentation when compared to the

control strains. In fact, the results show

that the production of the weapons

occurs independently of acetate

catabolism, in opposition to original

background info2. The weapons were only affected when RNAIII in the agr loci was targeted, in

opposition with the original hypothesis2. Thus, a new double agent is brought to light.

There are various differentvarious genes that have been found to affect the activity of aconitase; these are

the agents responsible. The first agent, called rsbU in the field, is responsible for positively regulating

sigma factor σB. With the downregulation caused by an 11-bp deletion, acetate catabolism is decreased.

The second agent, referred to as sdhB, had a single base pair deletion, altering S. aureus ability to encode

succinate dehydrogenase. This results in the truncation of protein formation.

3:Somerville GA, Chaussee MS, Morgan CI, Fitzgerald JR, Dorward DW, Reitzer LJ, Musser JM. 2002. Staphylococcus aureus Aconitase
Inactivation Unexpectedly Inhibits Post-Exponential-Phase Growth and Enhances Stationary-Phase Survival. Infect and Immun. 70(11):6373-
6382. DOI: 10.1128/iai.70.11.6373-6382.2002
Image 2: https://www.researchgate.net/figure/Schematic-map-of-the-S-aureus-agr-locus-showing-the-locations-of-the-different-
primers_fig1_259477171
 3

There are also three deactivated mobsters in the

aforementioned strainsstrains of S. aureus: citB/acnA,

odhA, and odhB. AcnA is a single aconitase gene 3,

essentially dictating the catabolism of acetate by

aconitase. CitB has a similar role as acnA, coding

aconitate hydratase A4. OdhA is responsible for coding a

subunit of the α-ketoglutarate dehydrogenase complex.

This felon was taken out with a 66-bp deletion, rendering

the subsequent S. aureus cells inableunable to catabolize

acetate. OdhB is used in catalyzing conversion to succinyl-CoA, relating to acetate 5. We are unaware at

this time which agent has been taking out opposition members, or what was used to inactive mobsters

citB/acnA and odhB.

In essence, pathogens are very similar to crime organizations; there’s always a double agent looking to

take you down. The crime headquatersheadquarters (pathogen cell) is filled with both mobsters (genes

helping) and agents (genes harming) alike. And Dr. Somerville and his team have been witnesswitnessing

to the whole scuffle.

Word count 737

10/15

4:Gao W, Dai S, Liu Q, Xu H, Bai Y, Qiao M. 2010. Effect of site-directed mutagenesis of citB on the expression and activity of Bacillus subtilis
aconitase. Microbiol. 79(6):774-778. URL:https://pubmed.ncbi.nlm.nih.gov/21446632/
5:Teoh WP, Resko ZJ, Flury S, Alonzo F. 2019. Dynamic Relay of Protein-Bound Lipoic Acid in Staphylococcus aureus. J Bacteriol.
201(22):e00446-19. DOI:10.1128/JB.00446-19
Image 3:Somerville GA, Said-Salim B, Wickman JM, Raffel SJ, Kreiswirth BN, Musser JM. 2003. Correlation of Acetate Catabolism and
Growth Yield in Staphylococcus aureus: Implications for Host-Pathogen Interactions. Infect and Immun. 71(8): 4724-4732. DOI:
10.1128/iai.71.8.4724-4732.2003