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Characterization of Mn(II) homeostasis within Bacillus subtilis
Author(s) -
Baertlein Matt,
Vos Cullen,
Jeffries Will,
Isaacson Erin,
Gabriel Scott
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1055.15
Subject(s) - efflux , bacillus subtilis , homeostasis , chemistry , metal , transcription factor , regulator , biochemistry , cofactor , enzyme , biology , microbiology and biotechnology , gene , bacteria , genetics , organic chemistry
Metal ion homeostasis is a critical component of all cellular life. Metals act as cofactors for many essential enzymes; yet, in excess, metals can cause oxidative stress and cell death. In order to further understand metal ion homeostasis, investigation of pathways that regulate the corresponding intake and efflux of metal ions is needed. While many of the pathways which control metal ion intake and efflux within B. subtilis have been characterized, Mn(II) efflux still remains unknown. Zone of inhibition (ZOI) data from our lab shows a ydfM knockout decreases the survivability of B. subtilis in the presence of toxic Mn(II) levels, while there was no significant difference in toxic concentrations of Cu(II), Cd(II), and Zn(II). Additionally, Beta‐galactosidase assays have shown that ydfM transcription increases with increasing Mn(II) concentration but not increasing concentrations of Cd(II), Cu(II), Fe(III), Ni(II), or Zn(II). These results have led us to hypothesize that that YdfM acts as an Mn(II) efflux protein within B. subtilis . Concurrently, we also have aimed to identify the transcriptional regulator of YdfM. Interestingly, the gene immediately downstream of YdfM is predicted to be a metal efflux protein regulator. Initial bioinformatics work, shows that YdfL possesses a helix turn helix DNA binding motif as well as several conserved amino acid sequences which suggest it is similar to other metal ion influx and efflux regulators. ZOI data also shows there is a significant increase in survivability in the ydfL knockout when compared to wild type B. subtilis , further suggesting YdfL acts as a regulatory repressor of ydfM . Additional preliminary beta‐galactosidase, growth curve, and gel shift data currently ongoing will also be presented which aims to confirm YdfM's role in the efflux of Mn(II) as well as YdfL's role as a regulator of ydfM transcription.