Binding of MAP3773c Protein of <em>Mycobacterium avium</em> subsp. <em>paratuberculosis </em>in the Mouse Ferroportin1 Coding Region

• DOI: 10.20944/preprints202410.1423.v1
• Publication Date: 2024-10-21T03:10:53Z
• Authors: Bertha Landeros S...; Dulce Liliana Due...; Jose Angel Gutier...; Kaina Quintero Ch...; Mirna Del Carmen ...; Dania Melissa Dia...; Omar Cortéz Herná...; José Román Chávez...; Jocelyn Marcela A...; Giselle Berenice ...

(1) Background: Mycobacterium avium subsp. paratuberculosis (MAP) is a bacterium that causes paratuberculosis disease. One of its proteins, MAP3773c, is involved in iron metabolism and acts as a transcription factor. Our goal is to investigate the binding of the MAP3773c protein in 2 sites of the chromatin of the ferroportin1 gene of mouse macrophages J774 A.1. (2) Methods: We performed a sequence alignment, then a docking, then EMSA to analyze in vitro the binding of MAP3773c, then we performed the cloning of map3773c in pCDNA3.1 to perform transfection in macrophages, then we performed the expression by Western blotting and PCR in real time then a CHiP of transfected macrophages. (3) Results: Our results showed in silico that MAP3773c binds to two boxes in the FPN1 coding sequence, by EMSA, MAP3773c is glued at 2 sites: EBOX and MBOX, equally with iron, zinc or EDTA. In CHiP, we also determined the binding of MAP3773c in the chromatin of FPN1 by PCR amplifying the chromatin fragment. (4) Conclusions: Our in silico, in vitro and in vivo study showed that MAP3773c binds to FPN1. Overall, our study provides information on the role of MAP3773c and its effect on host iron transport.