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1.E.53 The Toxic Hok/Gef Protein (Hok/Gef) Family

Members of the Hok/Gef family include a number of small proteins (about 50 amino acid residues), including the highly similar Gef and RelA proteins of E. coli and several plasmid-encoded proteins such as Hok, FimA, SrnB and PndA (Poulsen et al. 1989). These cell-toxic proteins have N-terminal hydrophobic ''signal'' peptides which traverse the cytoplasmic membrane of the bacteria displaying their C-terminal domains in the periplasm (Poulsen et al. 1991). When overexpressed, these proteins are toxic, killing the bacterial cell from the inside, but their normal physiological functions were not known. They appear to mediate programmed cell death in bacteria, and they function in plasmid maintenance by killing plasmid-free cells via a toxin/unstable (protease-sensitive) antitoxin-dependent mechanism. Members of the Hok/Gef family are found in bacteria and archaea, and the E. coli chromosome encodes at least five hok gene paralogues. Gef forms disulfide-linked homodimers. It has been proposed that they oligomerize to form transmembrane channels or pores that dissipate the membrane potential and thereby kill the cell.  This conclusion has been substantiated (Brielle et al. 2016).

Hok-like proteins are very toxic to most Gram-negative species and also to some extent to Gram-positive bacteria (Gerdes et al. 1997). Induction of Hok leads to loss of the cell membrane potential, arrest of respiration, efflux of small molecules (i.e. Mg2+ and ATP), influx of small extracellular molecules (i.e. ONPG) and even influx of periplasmic proteins such as RNase I. By phase contrast microscopy, the cells change morphology to so-called ''ghost-cells'' after induction of Hok protein synthesis. These ghost cells are characterized by condensed cell poles and a centrally located clearing, and resemble the ghost cells (empty cell shells) formed after induction of the lysis gene of φx174. Furthermore, the srnB gene of F complements mutations in the φx174 BS gene , which encodes a holin. Thus the Hok family of proteins bears functional resemblance to holins (TC subclass 1.E). The holins create holes or pores in the inner cell membrane, thereby releasing a phage-encoded endolysin to the periplasm, where the enzyme degrades the cell wall. Thus, the Hok-like proteins kill the cells by mediating irreversible damage to the host cell membrane.

References associated with 1.E.53 family:

Boulaiz, H., J. Prados, J.A. Marchal, A.M. García, L. Alvarez, C. Melguizo, E. Carrillo, J.L. Ramos, and A. Aránega. (2003). Transfection of MS-36 melanoma cells with gef gene inhibits proliferation and induces modulation of the cell cycle. Cancer Sci 94: 564-568. 12824883
Brielle, R., M.L. Pinel-Marie, and B. Felden. (2016). Linking bacterial type I toxins with their actions. Curr. Opin. Microbiol. 30: 114-121. [Epub: Ahead of Print] 26874964
Gerdes, K., A.P. Gultyaev, T. Franch, K. Pedersen, and N.D. Mikkelsen. (1997). Antisense RNA-regulated programmed cell death. Annu Rev Genet 31: 1-31. 9442888
Harms, A., E. Maisonneuve, and K. Gerdes. (2016). Mechanisms of bacterial persistence during stress and antibiotic exposure. Science 354:. 27980159
Poulsen, L.K., A. Refn, S. Molin, and P. Andersson. (1991). Topographic analysis of the toxic Gef protein from Escherichia coli. Mol. Microbiol. 5: 1627-1637. 1943700
Poulsen, L.K., N.W. Larsen, S. Molin, and P. Andersson. (1989). A family of genes encoding a cell-killing function may be conserved in all gram-negative bacteria. Mol. Microbiol. 3: 1463-1472. 2693900
Verstraeten, N., W.J. Knapen, C.I. Kint, V. Liebens, B. Van den Bergh, L. Dewachter, J.E. Michiels, Q. Fu, C.C. David, A.C. Fierro, K. Marchal, J. Beirlant, W. Versées, J. Hofkens, M. Jansen, M. Fauvart, and J. Michiels. (2015). Obg and Membrane Depolarization Are Part of a Microbial Bet-Hedging Strategy that Leads to Antibiotic Tolerance. Mol. Cell 59: 9-21. 26051177
Wilmaerts, D., M. Bayoumi, L. Dewachter, W. Knapen, J.T. Mika, J. Hofkens, P. Dedecker, G. Maglia, N. Verstraeten, and J. Michiels. (2018). The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage. MBio 9:. 30108166