The MinD Membrane Targeting Sequence Is a Transplantable Lipid-binding Helix

MinD is a ubiquitous ATPase that plays a crucial role in selection of the division site in eubacteria, chloroplasts, and probably also Archaea. It was recently demonstrated that membrane localization of MinD is mediated by an 8–12-residue C-terminal motif termed the membrane targeting sequence or MTS. In this study we show that the MinD MTS is a transplantable lipid-binding motif that can effectively target heterologous proteins to the cell membrane. We demonstrate that eubacterial MTSs interact directly with lipid bilayers as an amphipathic helix, with a distinct preference for anionic phospholipids. Moreover, we provide evidence that the phospholipid preference of each MTS, as well as its affinity for biological membranes, has been evolutionarily “tuned” to its specific role in different bacteria. We propose a model to describe how the MTS is coupled to ATP binding to regulate the reversible membrane association of Escherichia coli MinD during its pole-to-pole oscillation cycle. MinD is a ubiquitous ATPase that plays a crucial role in selection of the division site in eubacteria, chloroplasts, and probably also Archaea. It was recently demonstrated that membrane localization of MinD is mediated by an 8–12-residue C-terminal motif termed the membrane targeting sequence or MTS. In this study we show that the MinD MTS is a transplantable lipid-binding motif that can effectively target heterologous proteins to the cell membrane. We demonstrate that eubacterial MTSs interact directly with lipid bilayers as an amphipathic helix, with a distinct preference for anionic phospholipids. Moreover, we provide evidence that the phospholipid preference of each MTS, as well as its affinity for biological membranes, has been evolutionarily “tuned” to its specific role in different bacteria. We propose a model to describe how the MTS is coupled to ATP binding to regulate the reversible membrane association of Escherichia coli MinD during its pole-to-pole oscillation cycle. The initiating event in bacterial cytokinesis is the formation of a circumferential ring of polymerized FtsZ, the ancestral homolog of eukaryotic tubulin (1Erickson H.P. Trends Cell Biol. 1997; 7: 362-367Abstract Full Text PDF PubMed Scopus (200) Google Scholar, 2Harry E.J. Mol. Microbiol. 2001; 40: 795-803Crossref PubMed Scopus (88) Google Scholar, 3Lutkenhaus J. Curr. Opin. Microbiol. 2002; 5: 548-552Crossref PubMed Scopus (52) Google Scholar). The FtsZ ring provides a scaffold onto which numerous proteins are subsequently assembled to form the functional division apparatus (4Buddelmeijer N. Beckwith J. Curr. Opin. Microbiol. 2002; 5: 553-557Crossref PubMed Scopus (115) Google Scholar, 5Errington J. Daniel R.A. Scheffers D.J. Microbiol. Mol. Biol. Rev. 2003; 67: 52-65Crossref PubMed Scopus (531) Google Scholar). In the rod-shaped bacterium Escherichia coli, placement of the FtsZ ring, and thus the division septum, is negatively regulated by the three proteins encoded by the minB operon: MinC, MinD, and MinE (3Lutkenhaus J. Curr. Opin. Microbiol. 2002; 5: 548-552Crossref PubMed Scopus (52) Google Scholar, 6de Boer P.A.J. Crossley R.E. Rothfield L.I. Cell. 1989; 56: 641-649Abstract Full Text PDF PubMed Scopus (618) Google Scholar, 7Rothfield L.I. Shih Y.-L. King G.F. Cell. 2001; 106: 13-16Abstract Full Text Full Text PDF PubMed Scopus (48) Google Scholar). In the absence of the Min system, FtsZ rings can form either at midcell or in the nucleoid-free regions at either of the cell poles (8Yu X.-C. Margolin W. Mol. Microbiol. 1999; 32: 315-326Crossref PubMed Scopus (219) Google Scholar). Polar divisions are nonproductive as they lead to the formation of chromosomeless minicells and multinucleate filaments. MinC and MinD associate to form an indiscriminate division inhibitor whose activity is restricted to polar sites in E. coli by the action of MinE (6de Boer P.A.J. Crossley R.E. Rothfield L.I. Cell. 1989; 56: 641-649Abstract Full Text PDF PubMed Scopus (618) Google Scholar). Studies of GFP 1The abbreviations used are: GFP, green fluorescent protein; BsMinD, Bacillus subtilis MinD; BsMTS, B. subtilis membrane targeting sequence; CL, cardiolipin; EcMinD, Escherichia coli MinD; EcMTS, E. coli membrane targeting sequence; MTS, membrane targeting sequence; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; SUV, small unilamellar vesicle; WT, wild-type; LZ, leucine zipper.-labeled Min proteins from the Gram-negative bacteria E. coli and Neisseria gonorrhoeae have revealed that they undergo a remarkable pole-to-pole oscillation that causes the time-averaged concentration of the MinCD division inhibitor to be lowest at midcell (9Raskin D.M. de Boer P.A.J. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 4971-4976Crossref PubMed Scopus (602) Google Scholar, 10Raskin D.M. de Boer P.A.J. J. Bacteriol. 1999; 181: 6419-6424Crossref PubMed Google Scholar, 11Hu Z. Lutkenhaus J. Mol. Microbiol. 1999; 34: 82-90Crossref PubMed Scopus (370) Google Scholar, 12Rowland S.L. Fu X. Sayed M.A. Zhang Y. Cook W.R. Rothfield L.I. J. Bacteriol. 2000; 182: 613-619Crossref PubMed Scopus (97) Google Scholar, 13Fu X. Shih Y.-L. Zhang Y. Rothfield L.I. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 981-985Google Scholar, 14Hale C.A. Meinhardt H. de Boer P.A. EMBO J. 2001; 20: 1563-1572Crossref PubMed Scopus (231) Google Scholar, 15Ramirez-Arcos S. Szeto J. Dillon J.-A. Margolin W. Mol. Microbiol. 2002; 46: 493-504Crossref PubMed Scopus (42) Google Scholar). This makes midcell the preferred site for construction of an FtsZ ring. In contrast, Bacillus subtilis and most other Gram-positive bacteria lack the MinE protein that promotes MinCD oscillation in E. coli. Instead, in B. subtilis, the MinCD complex is anchored at the cell poles by DivIVA where it remains throughout the cell cycle until a late stage in assembly of the division apparatus when it is piloted to the nascent division site (16Marston A.L. Thomaides H.B. Edwards D.H. Sharpe M.E. Errington J. Genes Dev. 1998; 12: 3419-3430Crossref PubMed Scopus (291) Google Scholar, 17Marston A.L. Errington J. Mol. Microbiol. 1999; 33: 84-96Crossref PubMed Scopus (158) Google Scholar, 18Harry E.J. Lewis P.J. Mol. Microbiol. 2003; 47: 37-48Crossref PubMed Scopus (50) Google Scholar). MinD is a peripheral membrane protein (19de Boer P.A.J. Crossley R.E. Hand A.R. Rothfield L.I. EMBO J. 1991; 10: 4371-4380Crossref PubMed Scopus (265) Google Scholar), and its association with the inner membrane is a prerequisite for subsequent membrane recruitment of MinC (and MinE in E. coli). MinD is a member of the ParA superfamily of ATPases that are characterized by a deviant Walker A motif (19de Boer P.A.J. Crossley R.E. Hand A.R. Rothfield L.I. EMBO J. 1991; 10: 4371-4380Crossref PubMed Scopus (265) Google Scholar, 20Yamaichi Y. Niki H. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 14656-14661Crossref PubMed Scopus (134) Google Scholar, 21Gerdes K. Møller-Jensen J. Jensen R.B. Mol. Microbiol. 2000; 37: 455-466Crossref PubMed Scopus (359) Google Scholar, 22Lutkenhaus J. Sundaramoorthy M. Mol. Microbiol. 2003; 48: 295-303Crossref PubMed Scopus (97) Google Scholar). The ATPase activity of MinD provides the driving force for oscillation of the Min proteins in E. coli; this activity is stimulated by MinE but only in the presence of phospholipids (23Hu Z. Lutkenhaus J. Mol. Cell. 2001; 7: 1337-1343Abstract Full Text Full Text PDF PubMed Scopus (217) Google Scholar, 24Hu Z. Gogol E.P. Lutkenhaus J. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar, K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google Scholar). In in the presence of ATP and phospholipid MinD and K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google as well as that to a of MinD Z. Gogol E.P. Lutkenhaus J. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). in E. coli MinD to be that the inner with each to of Y.-L. Rothfield Proc. Natl. Acad. Sci. U. S. A. 2003; PubMed Scopus Google Scholar). The by which MinD with the inner membrane for a it was recently demonstrated that MinD is to the membrane by a C-terminal motif S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar, Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google that we to as the membrane targeting sequence or MTS. We that this which is a amphipathic that a MinD and membrane phospholipids S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). this a of the MTS biological with different phospholipid can the MTS membrane association of B. subtilis MinD but for of membrane in the of E. coli MinD the MTS as an In this we show that the MinD MTS directly with lipid bilayers as an amphipathic helix, and that it can as a transplantable Moreover, we provide evidence that the phospholipid preference and membrane affinity of each MTS has been “tuned” to its specific role in different bacteria. We propose a model to describe how the MTS is coupled to ATP binding to regulate the reversible membrane association of during its oscillation cycle. to of with an of and with a sequence by The of to the of the MinD The of the was to of a that be to formation E.J. PubMed Scopus Google Scholar). by The concentration of each for was by of and and E. coli from Polar as M.E. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google with in at the in and The in to a concentration of for in a until the used for the they at a and lipid in in a The was at the of a of and a of only or lipid was from the as The of each was from the to the PubMed Scopus Google Scholar, PubMed Scopus Google Scholar), where is the of the is the of the at is the at of an of is the of to be is a with a of at and is the of in the was from of proteins in it has been used to the of to lipid bilayers M.E. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, R.B. 33: PubMed Scopus Google Scholar, R.B. PubMed Scopus Google Scholar, H. 1999; PubMed Scopus Google Scholar). of GFP GFP used in this study of the and with We used the for GFP S. PubMed Scopus Google Scholar). A GFP to was by S.L. Fu X. Sayed M.A. Zhang Y. Cook W.R. Rothfield L.I. J. Bacteriol. 2000; 182: 613-619Crossref PubMed Scopus (97) Google Scholar), a as the and that This the of from and by to GFP and of the as was used as a in a with and to a that encoded the This was with and a that GFP at the to the to is was by the with and it to This a of the to the of GFP a C-terminal sequence is to the protein encoded by of the as the and GFP to and of with each by a was by the B. subtilis as S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google and and and This was an to in which is encoded as an to the of GFP was as was used to with and and and The to to the of A was and to for A was used to and the leucine of the was by a a as King G.F. Sci. 1999; Scholar). The and This with a and was as of GFP was as S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google the E. coli (6de Boer P.A.J. Crossley R.E. Rothfield L.I. Cell. 1989; 56: 641-649Abstract Full Text PDF PubMed Scopus (618) Google that a of the minB The MinD MTS with recently that the MinD MTS directly with as an amphipathic that the the the polar of the interact with the of anionic phospholipids S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). this we used to the of the MTS of E. coli and B. subtilis MinD to interact directly with lipid bilayers in the form of In the absence of the and with a at that is of a A and in the absence of lipid the E. coli the B. subtilis MTS This is with the that the C-terminal of MinD are when the protein is in the absence of J. 2001; PubMed Scopus Google Scholar). We the of MTS with a of PE, PG, that the phospholipid in the E. coli inner membrane W. 1998; 455-466Crossref PubMed Scopus Google Scholar, B. Curr. 1997; Scopus Google we to as of to the a in the the at was to and the at This that the directly with lipid and the of the at that this causes the MTS to at in the presence of phospholipid bilayers M. J. J. Mol. Biol. 1999; PubMed Scopus Google have been for the of the M.E. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google and the E. coli A. J. Biol. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The at that the which to We that to which the of the MTS by a in in to the membrane S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). A this is in the absence of and in the presence of We that the amphipathic of the MTS is for its with of to the a in the with the the at was to and the at The from the at is this to which is the of the B. subtilis MTS S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). A a of the sequence was in the absence of and in the presence of This that the specific sequence of in the MTS, and the of the is for membrane We that the E. coli and B. subtilis MTS interact directly with phospholipid bilayers as amphipathic The be only an of the of each MTS the in the of the and in the presence of is as by the in of the at in the of the with are of this in it a in the of the by the and when they lipid We this is as the of the when to is only to and we are of a small and lipid The that we is that the by the and are of and that the for the that it biological with affinity the The is an the of the and it the and of the from the and we that the and form in the but the has affinity for the EcMTS, the of be the of the and it to be as we other are we the when the MTS with as an that the has affinity for phospholipid MTS for the of the polar of the MTS in A and we that the E. coli and B. subtilis MTSs interact with negatively phospholipids as and S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). with this it was recently demonstrated that to anionic phospholipids E. Fu X. B. Margolin W. W. J. Biol. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar). this of MinD in the MTS, we the of the and to with a of but of the anionic phospholipids and we anionic phospholipids are for MTS binding by with the phospholipid In to the with the of the and was by the of of the phospholipids and and green in A and In other interact with We that the E. coli and B. subtilis MTS can only interact with biological anionic phospholipids. the different phospholipid of the cell of E. coli and B. subtilis, we MinD small with different anionic phospholipids. the a distinct preference for This was demonstrated by the that PE, a for this of PE, the and in The from the at was for the in the presence of in the presence of We that the with biological In to the with the B. subtilis MTS, the well with and The by of was different to the by that a of and We that the with bilayers anionic but it has preference for or The MinD MTS a that membrane localization of and in an E. coli was the MTS S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). of as as three C-terminal in the of or in the of BsMinD, as well as of the of the MTS helix, was to MinD localization S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). This that the MTS is but for membrane localization of It was recently that GFP was to the membrane when the C-terminal of to its or Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google Scholar), that other regions of also be in its association with the inner membrane. the MinD MTS as a transplantable membrane targeting we the of the and to target GFP to the membrane of the E. coli We that GFP is in the of S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). when the C-terminal of to the of GFP, the protein was to the cell of membrane association We also the target to the cell membrane. MinC is the inhibitor of E. coli cytokinesis Boer P.A.J. Crossley R.E. Rothfield L.I. J. Bacteriol. PubMed Scopus Google but it is only when to the membrane by MinD Z. Lutkenhaus J. J. Bacteriol. 2003; PubMed Scopus Google Scholar). was to be in E. coli D.M. de Boer P.A.J. J. Bacteriol. 1999; 181: 6419-6424Crossref PubMed Google Scholar), we that a protein was to the cell in this the is of targeting proteins of at the of the protein to the cell membrane. We that the is an and transplantable membrane targeting a a of and of of the minB (6de Boer P.A.J. Crossley R.E. Rothfield L.I. Cell. 1989; 56: 641-649Abstract Full Text PDF PubMed Scopus (618) Google Scholar), we that of the to of a to recruitment of to the membrane the causes the cell division of MinC to in the absence of It was recently in to MinC to the MinC to nascent de Boer P.A. J. Bacteriol. 2002; PubMed Scopus Google Scholar). the that MinC when to the membrane in a that targeting of MinC to is for of its In to the with BsMTS, GFP in the when the C-terminal of to its This from the that the has a affinity for phospholipid bilayers the This the of how remains to membrane to ATP The in which the was to GFP that a provide affinity for to lipid a that MinD an to the that the MTS have affinity for the membrane Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google Scholar), binding of the MTS to the membrane effectively be an We this in we the localization of a in which a leucine motif was the of GFP and of this GFP we used an of the leucine that was to form the sequence King G.F. Sci. 1999; Scholar). A was the leucine and to that the of the MTS was by the leucine GFP when with the leucine motif but the GFP to the membrane In a we the of GFP with a of or of the and the to the cell We that a but a can associate with the E. coli cell membrane. the peripheral localization was as for the as we with the This that the affinity of the only the of the EcMTS, but also the of the MTSs in The of MinD was recently that membrane localization of MinD a C-terminal MTS that is eubacteria, and S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar, Z. Lutkenhaus J. J. Bacteriol. 2003; PubMed Scopus Google Scholar). In this we the S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google that the MTS directly with membrane phospholipids as an amphipathic helix, with a distinct preference for anionic phospholipids. Moreover, we demonstrated that the MTSs from different bacteria interact with different of anionic a remarkable the small of this motif This makes the membrane of bacteria can we that the with anionic it and PG, with the that is only in the E. coli inner membrane W. 1998; 455-466Crossref PubMed Scopus Google Scholar, B. Curr. 1997; Scopus Google Scholar). In contrast, we that the has a distinct preference for CL, with the that are of of and but in B. subtilis cell K. M. Y. H. M. J. Bacteriol. 1998; PubMed Google Scholar). the polar of each MTS to have been evolutionarily for with specific phospholipids that are in the inner membrane of the bacterium in which it In this it be in to the lipid of the from whose have a different lipid to H. N. Y. A. J. Bacteriol. 2002; PubMed Scopus Google Scholar). of the of an complex probably be for the of MTS lipid MTS for a MTS from to be that from is The in the membrane of the which in is to the different localization and biological of E. coli and B. subtilis it remains anchored at the cell poles by DivIVA until it is to the nascent at a late stage in assembly of the division (16Marston A.L. Thomaides H.B. Edwards D.H. Sharpe M.E. Errington J. Genes Dev. 1998; 12: 3419-3430Crossref PubMed Scopus (291) Google Scholar, 18Harry E.J. Lewis P.J. Mol. Microbiol. 2003; 47: 37-48Crossref PubMed Scopus (50) Google Scholar). to to the cell membrane throughout the cell cycle. We that the has been to have affinity for phospholipid bilayers to the association of with the cell membrane. demonstrated in this the affinity of the for lipid bilayers it to as a transplantable membrane targeting motif that to be a for proteins to the cell membrane. In to the B. subtilis a pole-to-pole oscillation that throughout the cell cycle. This oscillation of membrane S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). ATP binding promotes association of MinD with the ATP to of MinD from the (23Hu Z. Lutkenhaus J. Mol. Cell. 2001; 7: 1337-1343Abstract Full Text Full Text PDF PubMed Scopus (217) Google Scholar, 24Hu Z. Gogol E.P. Lutkenhaus J. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar, K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google Scholar, Z. Lutkenhaus J. J. Bacteriol. 2003; PubMed Scopus Google Scholar). We that an MTS with affinity for the be to we that the is for membrane affinity that can be by ATP at the in the oscillation cycle. A for the of is the that the reversible association of the with the of the inner We with the J. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar), that the of the MTS with the of MinD when the protein is in the or thus association with the membrane. In this ATP binding a in MinD that the MTS and promotes association with the S.L. Rothfield L.I. King G.F. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus Google Scholar). In a of this it was that the be by of MinD, which in of a MTS with affinity for the membrane J. Sundaramoorthy M. Mol. Microbiol. 2003; 48: 295-303Crossref PubMed Scopus (97) Google Scholar, Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google Scholar). are with they are to with the that the C-terminal of A. MinD, which the MTS, are when the protein is in the absence of and J. 2001; PubMed Scopus Google that the model in which membrane association is by a the MTS and the of has been for E. coli MinD in Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google Scholar, Z. Lutkenhaus J. J. Bacteriol. 2003; PubMed Scopus Google Scholar), it S. and Moreover, that the of N. gonorrhoeae MinD be J. S. Dillon J. Bacteriol. 2001; PubMed Scopus (52) Google Scholar), which that regulate membrane of MinD in this We propose an for the reversible membrane association of that of In this which we to as the or model the and of MinD are and a MTS that has only affinity for the membrane. the and of MinD be in the membrane and but the be the in this the MTS be in the of MinD and in the The of this model is that promotes of MinD the membrane. association of the form of MinD with the membrane of the protein that a MTS a MTS. The MTS thus a that and to the membrane as MinD and thus the MinD of a MTS with membrane affinity a MTS with membrane The MinD is to at of the cell poles and midcell Y.-L. Rothfield Proc. Natl. Acad. Sci. U. S. A. 2003; PubMed Scopus Google Scholar, Fu X. King G.F. Rothfield L.I. EMBO J. 2002; PubMed Scopus Google Scholar). MinE probably MinD in at We that MinE the or of the to This is to the role of in FtsZ by E.J. J. Proc. Natl. Acad. Sci. U. S. A. 2003; PubMed Scopus Google Scholar). The absence of MinE rings and the MinD polar in a MinE Fu X. King G.F. Rothfield L.I. EMBO J. 2002; PubMed Scopus Google that this be in the binding of MinE at the of the MinD is to ATP at the of the MinD (23Hu Z. Lutkenhaus J. Mol. Cell. 2001; 7: 1337-1343Abstract Full Text Full Text PDF PubMed Scopus (217) Google Scholar, K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google in the this the MinD which is in the and the of the the MinD at the of the from the membrane the form of the protein is to and its affinity for the membrane in a MTS. In contrast, the of the MinD remains by the MTS for is MinE to at the of the MinD We that the MinD eukaryotic and is that MinE can only to the It is well that proteins to the of J. 2003; PubMed Scopus Google and that proteins and to the of Curr. Opin. Cell Biol. 2000; 12: PubMed Scopus Google Scholar). evidence that the MinD is polar from in that MinE causes at only of MinD K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google Scholar). The model is by of a of have demonstrated that lipid bilayers with (23Hu Z. Lutkenhaus J. Mol. Cell. 2001; 7: 1337-1343Abstract Full Text Full Text PDF PubMed Scopus (217) Google Scholar, E. Fu X. B. Margolin W. W. J. Biol. 2003; Full Text Full Text PDF PubMed Scopus Google as by the in with the of A. MinD, the model the MTS to be from the membrane or in in the and of MinD; membrane association the of the MTS that in is by the of MinD as demonstrated in this and are to target GFP to the E. coli a is as by the MTS to membrane The of has been to that with the membrane as a Z. Lutkenhaus J. Mol. Microbiol. 2003; 47: PubMed Scopus Google Scholar, Z. Lutkenhaus J. J. Bacteriol. 2003; PubMed Scopus Google Scholar). the that in in the absence of to form K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: PubMed Scopus (134) Google that most the assembly of MinD the membrane. the of the model the of the MinD is a or it be in to is an prerequisite for of MinD the membrane. We for and Dillon for and for in the of this