Identification of human alpha?enolase as an interaction partner of a Brucella abortus Virb substrate

MORRONE SEIJO S.; MARCHESINI M.I; GUAIMAS F.; COMERCI D. J

Rosario

Congreso; L Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2014

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Brucella abortus, the causative agent of bovine brucellosis, invades and replicates within cellsinside a membrane? bound compartment known as the Brucella Containing Vacuole (BCV).Brucella Type IV Secretion System (VirB) translocates effector proteins across the BCV thoughtto modulate host cell signaling pathways to favor intracellular survival and replication. BPE123is a VirB?translocated effector protein recently identified. It is a hypothetical protein whosefunction remains unknown. In an attempt to identify host cell proteins interacting with BPE123,a pull? down assay was performed and human alpha?enolase (ENO1) was identified by LC/MSMSas a potential interaction partner of BPE123. These results were confirmed byimmunoprecipitation. Microscopy studies revealed ENO1 BIOCELL 38 (Suppl. 2) 2014 129relocalization upon ectopic expression of BPE123 in HeLa cells, where both proteins localized tothe ER. Furthermore, ENO1 is recruited to the vicinities of BPE123 positive BCVs in infectedmacrophages, indicating that interaction with translocated BPE123 might also be occurringduring the intracellular phase of B. abortus. These preliminary results suggest a direct interactionbetween BPE123 and ENO1, a multifunctional protein that is required for Brucella replication inhost cells. Further experiments are underway to determine how BPE123?ENO1 interactionmodulates the outcome of the infection process.