Role of MinD-membrane association in Min protein interactions.
Division site placement in Escherichia coli involves interactions of the MinD protein with MinC and MinE and with other MinD molecules to form membrane-associated polymeric structures. In this work, as part of a study of these interactions, we established that heterologous membrane-associated proteins such as MinD can be targeted to the yeast nuclear membrane, dependent only on the presence of a membrane-binding domain and a nuclear targeting sequence. Targeting to the nuclear membrane was equally effective using the intrinsic MinD membrane-targeting domain or the completely unrelated membrane-targeting domain of cytochrome b(5). The chimeric proteins differing in their membrane-targeting sequences were then used to establish the roles of membrane association and specificity of the membrane anchor in MinD interactions, using the yeast two-hybrid system. The chimeric proteins were also used to show that the membrane association of MinD and MinE in E. coli cells had no specificity for the membrane anchor, whereas formation of MinDE polar zones and MinE rings required the presence of the native MinD membrane-targeting sequence.
Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, Farmington, CT 06032, USA. taghbalout@neuron.uchc.edu
Aziz
A
Taghbalout
Luyan
L
Ma
Lawrence
L
Rothfield
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D018797:Cell Cycle Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D015786:Cytochromes b5 / Q000235:genetics; D003937:Diagnosis, Differential; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008958:Models, Molecular; D009685:Nuclear Envelope / Q000737:chemistry; D011485:Protein Binding; D021382:Protein Sorting Signals / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D021381:Protein Transport / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C485182:Cyan Fluorescent Protein; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; C093548:MinE protein, E coli; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D015786:Cytochromes b5; D000251:Adenosine Triphosphatases; C072069:MinD protein, E coli