title: R174 of Escherichia coli FtsZ is involved in membrane interaction and protofilament bundling, and is essential for cell division.
abstract: We investigated the interaction between FtsZ and the cytoplasmic membrane using inside-out vesicles. Comparison of the trypsin accessibility of purified FtsZ and cytoplasmic membrane-bound FtsZ revealed that the protruding loop between helix 6 and helix 7 is protected from trypsin digestion in the latter. This hydrophobic loop contains an arginine residue at position 174. To investigate the role of R174, this residue was replaced by an aspartic acid, and FtsZ-R174D was fused to green fluorescent protein (GFP). FtsZ-R174D-GFP could localize in an FtsZ and in an FtsZ84(Ts) background at both the permissive and the non-permissive temperature, and it had a reduced affinity for the cytoplasmic membrane compared with wild-type FtsZ. FtsZ-R174D could also localize in an FtsZ depletion strain. However, in contrast to wild-type FtsZ, FtsZ-R174D was not able to complement the ftsZ84 mutation or the depletion strain and induced filamentation. In vitro polymerization experiments showed that FtsZ-R174D is able to polymerize, but that these polymers cannot form bundles in the presence of 10 mM CaCl2. This is the first description of an FtsZ mutant that has reduced affinity for the cytoplasmic membrane and does not support cell division, but is still able to localize. The mutant is able to form protofilaments in vitro but fails to bundle. It suggests that neither membrane interaction nor bundling is a requirement for initiation of cell division.
authors.affiliation: Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, Molecular Cytology, University of Amsterdam, Kruislaan 316, 1098 SM Amsterdam, The Netherlands.
authors.forename: Cecile-Marie
authors.initials: CM
authors.lastname: Koppelman
authors.forename: Mirjam E G
authors.initials: ME
authors.lastname: Aarsman
authors.forename: Jarne
authors.initials: J
authors.lastname: Postmus
authors.forename: Evelien
authors.initials: E
authors.lastname: Pas
authors.forename: Anton O
authors.initials: AO
authors.lastname: Muijsers
authors.forename: Dirk-Jan
authors.initials: DJ
authors.lastname: Scheffers
authors.forename: Nanne
authors.initials: N
authors.lastname: Nanninga
authors.forename: Tanneke
authors.initials: T
authors.lastname: den Blaauwen
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005816:Genetic Complementation Test; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011108:Polymers / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013696:Temperature; D014357:Trypsin / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; C076835:FtsZ84 protein, E coli; D011108:Polymers; D011993:Recombinant Fusion Proteins; D001120:Arginine; D014357:Trypsin