Analysis of MinD Mutations Reveals Residues Required for MinE Stimulation of the MinD ATPase and Residues Required for MinC Interaction

ABSTRACT The MinD ATPase is critical to the oscillation of the Min proteins, which limits formation of the Z ring to midcell. In the presence of ATP, MinD binds to the membrane and recruits MinC, forming a complex that can destabilize the cytokinetic Z ring. MinE, which is also recruited to the membrane by MinD, displaces MinC and stimulates the MinD ATPase, resulting in the oscillation of the Min proteins. In this study we have investigated the role of lysine 11, present in the deviant Walker A motif of MinD, and the three residues in helix 7 (E146, S148, and D152) that interact electrostatically with lysine 11. Lysine 11 is required for interaction of MinD with the membrane, MinC, MinE, and itself. In contrast, the three residues in helix 7 that interact with lysine 11 are not required for binding to the membrane or activation of MinC. They are also not required for MinE binding; however, they are required for MinE to stimulate the MinD ATPase. Interestingly, the D152A mutant self-interacts, binds to the membrane, and recruits MinC and MinE in the presence of ADP as well as ATP. This mutant provides evidence that dimerization of MinD is sufficient for MinD to bind the membrane and recruit its partners.

[1]  P. D. de Boer,et al.  MinDE-Dependent Pole-to-Pole Oscillation of Division Inhibitor MinC in Escherichia coli , 1999, Journal of bacteriology.

[2]  L. Rothfield,et al.  A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli , 1989, Cell.

[3]  Karsten Kruse,et al.  A dynamic model for determining the middle of Escherichia coli. , 2002, Biophysical journal.

[4]  J. Lutkenhaus,et al.  Topological regulation of cell division in Escherichia coli involves rapid pole to pole oscillation of the division inhibitor MinC under the control of MinD and MinE , 1999, Molecular microbiology.

[5]  J. Lutkenhaus,et al.  The MinC component of the division site selection system in Escherichia coli interacts with FtsZ to prevent polymerization. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Löwe,et al.  Crystal structure of the bacterial cell division regulator MinD , 2001, FEBS letters.

[7]  J. Lutkenhaus,et al.  Membrane Binding by MinD Involves Insertion of Hydrophobic Residues within the C-Terminal Amphipathic Helix into the Bilayer , 2003, Journal of bacteriology.

[8]  D. Belin,et al.  A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. , 1984, Gene.

[9]  L. Rothfield,et al.  The MinE ring required for proper placement of the division site is a mobile structure that changes its cellular location during the Escherichia coli division cycle. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Lutkenhaus,et al.  Analysis of MinC Reveals Two Independent Domains Involved in Interaction with MinD and FtsZ , 2000, Journal of bacteriology.

[11]  P. D. de Boer,et al.  Targeting of DMinC/MinD and DMinC/DicB Complexes to Septal Rings in Escherichia coli Suggests a Multistep Mechanism for MinC-Mediated Destruction of Nascent FtsZ Rings , 2002, Journal of bacteriology.

[12]  C. Touriol,et al.  Deletion analysis of gene minE which encodes the topological specificity factor of cell division in Escherichia coli , 1995, Molecular microbiology.

[13]  D. Belin,et al.  Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter , 1995, Journal of bacteriology.

[14]  P A de Boer,et al.  Dynamic localization cycle of the cell division regulator MinE in Escherichia coli , 2001, The EMBO journal.

[15]  D. Bramhill,et al.  Bacterial cell division. , 1997, Annual review of cell and developmental biology.

[16]  Yu-Ling Shih,et al.  Division site selection in Escherichia coli involves dynamic redistribution of Min proteins within coiled structures that extend between the two cell poles , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[17]  D. Rees,et al.  Structure of ADP·AIF4 –-stabilized nitrogenase complex and its implications for signal transduction , 1997, Nature.

[18]  P. D. de Boer,et al.  ATP-Dependent Interactions between Escherichia coli Min Proteins and the Phospholipid Membrane In Vitro , 2003, Journal of bacteriology.

[19]  K. Morikawa,et al.  Structural and functional studies of MinD ATPase: implications for the molecular recognition of the bacterial cell division apparatus , 2001, The EMBO journal.

[20]  P A de Boer,et al.  Rapid pole-to-pole oscillation of a protein required for directing division to the middle of Escherichia coli. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Lutkenhaus,et al.  A conserved sequence at the C‐terminus of MinD is required for binding to the membrane and targeting MinC to the septum , 2003, Molecular microbiology.

[22]  N. Wingreen,et al.  Dynamic structures in Escherichia coli: Spontaneous formation of MinE rings and MinD polar zones , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[23]  L. Rothfield,et al.  The MinD protein is a membrane ATPase required for the correct placement of the Escherichia coli division site. , 1991, The EMBO journal.

[24]  J. Lutkenhaus,et al.  MinD and role of the deviant Walker A motif, dimerization and membrane binding in oscillation , 2003, Molecular microbiology.

[25]  C. Sáez,et al.  Recruitment of MinC, an Inhibitor of Z-Ring Formation, to the Membrane in Escherichia coli: Role of MinD and MinE , 2003, Journal of bacteriology.

[26]  Detlef D. Leipe,et al.  Classification and evolution of P-loop GTPases and related ATPases. , 2002, Journal of molecular biology.

[27]  D. Raskin,et al.  The MinE Ring: An FtsZ-Independent Cell Structure Required for Selection of the Correct Division Site in E. coli , 1997, Cell.

[28]  J. Lutkenhaus,et al.  Topological regulation of cell division in E. coli. spatiotemporal oscillation of MinD requires stimulation of its ATPase by MinE and phospholipid. , 2001, Molecular cell.

[29]  H. Meinhardt,et al.  Pattern formation in Escherichia coli: A model for the pole-to-pole oscillations of Min proteins and the localization of the division site , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Valluzzi,et al.  Dynamic assembly of MinD into filament bundles modulated by ATP, phospholipids, and MinE , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[31]  W. Margolin,et al.  Conservation of dynamic localization among MinD and MinE orthologues: oscillation of Neisseria gonorrhoeae proteins in Escherichia coli , 2002, Molecular microbiology.

[32]  William Dowhan,et al.  Effects of Phospholipid Composition on MinD-Membrane Interactions in Vitro and in Vivo* , 2003, Journal of Biological Chemistry.

[33]  E. Bi,et al.  Cell division inhibitors SulA and MinCD prevent formation of the FtsZ ring , 1993, Journal of bacteriology.

[34]  G. King,et al.  The MinD Membrane Targeting Sequence Is a Transplantable Lipid-binding Helix* , 2003, Journal of Biological Chemistry.

[35]  J. Lutkenhaus,et al.  Dynamic assembly of MinD on phospholipid vesicles regulated by ATP and MinE , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Lutkenhaus,et al.  FtsK is an essential cell division protein that is localized to the septum and induced as part of the SOS response , 1998, Molecular microbiology.

[37]  G. King,et al.  Membrane localization of MinD is mediated by a C-terminal motif that is conserved across eubacteria, archaea, and chloroplasts , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Beckwith,et al.  Septal Localization of FtsQ, an Essential Cell Division Protein in Escherichia coli , 1999, Journal of bacteriology.

[39]  J. Lutkenhaus,et al.  The Switch I and II Regions of MinD Are Required for Binding and Activating MinC , 2004, Journal of bacteriology.