GREEN FLUORESCENT PROTEIN TRANSFORMATION OF YERSINIA ENTEROCOLITICA SEROTYPE O:3

VELAZQUEZ, MARSOL; MANZUR, MARÍA JIMENA; SILVA JUAN; ELICABE, JAVIER; DI GENARO SILVIA

San Luis

Congreso; XXXVII Reunión científica anual-SOBIO 2019; 2019

Yersinia enterocolitica  (Ye) is a Gram-negative bacteria that cause gastrointestinal infection and eventually, extra intestinal manifestations. Ye evade the immune response by injecting Yersinia outer proteins (Yops) into the cytosol of host cells using type IIIsecretion system. In addition, the lectin Galectin-1 (Gal-1) is a proto-type member from the family of galectins widely distributed in immune compartments. Knowledge about the interconnection between Yops and endogen host proteins is limited.   The aim of thiswork was characterize the interaction between Gal-1 and Yops and obtain preliminary ?in silico? models of non crystallized virulence factors of Ye. Our results showed the presence of Gal-1-permissive glycoepitopes in the Yops. Moreover, we demonstrated that Gal-1 binds to Yops by glycan-protein interactions and that Gal-1 protects some Yops of trypsine digestion.  Additionally, the sequence corresponding to the virulence factor, YopE was obtained from the National Center for Biotechnology Information database (NCBI)and comparative modeling was performed by searching for resolved proteins structures in the Protein Data Bank (PDB).  We used the catalytic domain of YopE from Y. pestis (1HY5), YopE of Y. pseudotuberculosis (1L2W) and the human Helicase RECQL4 (5LST)as query. The results provided the potential templates for different primary structural regions. The alignments were used to perform the YopE homology modeling using MODELLER 9v12 software. The preliminary models obtained were evaluated using PyMOL and COOT software. In addition, the intrinsic disorder was assayed using DisEMBL y DisProt software. Future studies are needed to understand cellular and molecular mechanisms to clarify the potential clinical relevance of our findings, focusing in the interactions between Gal-1 and Yops during Ye infection.