Dissertations and Theses (Open Access)

Author ORCID Identifier

https://orcid.org/0009-0001-6539-419X

Date of Graduation

8-2026

Document Type

Dissertation (PhD)

Program Affiliation

Microbiology and Infectious Diseases

Degree Name

Doctor of Philosophy (PhD)

Advisor/Committee Chair

Anthony R. Flores, MD, MPH, PhD

Committee Member

Jennifer Walker, PhD

Committee Member

Samuel A. Shelburne, MD, PhD

Committee Member

Blake Hanson, PhD

Committee Member

Kathryn Patras, PhD

Abstract

Group A Streptococcus (GAS) is a human-restricted pathogen whose global incidence has surged in the post-COVID era. The ability of GAS to shift from a colonizing to invasive phenotype depends on coordinated virulence gene regulation in response to host-imposed stressors. The paralogous global transcriptional regulators, SpxA1 and SpxA2, have emerged as central mediators of these adaptive responses; however, the mechanisms by which these regulators influence both global gene regulation in direct response to external stresses and the host response as a result of regulatory changes remain incompletely understood. Using an integrated multi-omic approach, we define the distinct yet interconnected regulatory programs of SpxA1 and SpxA2 across multiple host-relevant stress conditions. RNA-seq revealed functionally distinct regulons, with SpxA1 governing oxidative stress defense and metal homeostasis and SpxA2 coordinating virulence-associated gene expression. Proteomic analysis identified SpxA2 as a ClpXP protease substrate and revealed reciprocal paralog accumulation upon loss of either regulator. NanoString profiling demonstrated that the ratio of SpxA1/SpxA2, rather than the activity of either paralog alone, determines which transcriptional programs are engaged. ChIP-exo established that SpxA2 directly modulates CovR-DNA binding occupancy, defining the LiaFSR-SpxA2-CovRS axis as a cross-regulatory circuit linking cell envelope stress sensing to virulence gene regulation. During macrophage infection, SpxA1 and SpxA2 exert divergent effects on intracellular survival and host cytokine production. Additionally, SpxA1 deletion impairs GAS growth under copper ion stress and dysregulates metal homeostasis pathways. Together, these findings establish SpxA1 and SpxA2 function as a regulatory rheostat that coordinate GAS adaptation to host-imposed pressures.

Keywords

transcriptional regulation, host-pathogen interactions, cell envelope stress, two-component systems, oxidative stress

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.