CHARACTERIZATION OF HOMO E. COLI, HETERO, AND CHIMERA AND ESKAPE GROES/GROEL, PARASITIC OLIGOMERIC T. BRUCEI HSP10 AND HSP60, AND THE MITOCHONDRIAL IMPORT SEQUENCE ON HUMAN HSP60

Abstract

The GroES/GroEL chaperonin system is essential to bacterial life. As such, it holds great potential as an antibiotic target. Previously, while investigating ESKAPE pathogen GroES/GroEL, we challenged the general assumption that E. coli GroES/GroEL accurately represents similar chaperones with high genetic similarity when several E. coli strains expressing ESKAPE pathogen GroES/GroEL were inviable. In this study, we built upon our prior investigation and generated viable E. coli strains expressing ESKAPE pathogen GroES/GroEL in GroES/GroEL-null E. coli. To determine the functional differences previously disrupting cellular vitality, we observed the formation of nonfunctional hetero-oligomeric P. aeruginosa- or E. faecium-E. coli tetradecameric GroEL and investigated GroEL domain incompatibilities impairing tetradecamer function. With viable E. coli strains expressing ESKAPE pathogen GroES/GroEL and a better understanding of GroEL incompatibilities affecting function, we expanded beyond bacteria and applied our investigation strategy to T. brucei and human mitochondrial HSP10/60 (eukaryotic GroES/GroEL homologs). We began generating viable E. coli strains expressing T. brucei HSP10/60 and human mitochondrial HSP10 and HSP60 or MIS-HSP60 to investigate the functional differences between E. coli GroES/GroEL and T. brucei HSP10/60 and the effect of the mitochondrial import sequence (MIS) on human mitochondrial HSP60 function.