Features of the plasmid pMV158-encoded MobM, a protein involved in its mobilization.
The streptococcal promiscuous plasmid pMV158 can be mobilized between a number of bacterial species by means of three elements: (i) the plasmid-encoded nicking-closing protein MobM, involved in the initiation and termination of the conjugative transfer; (ii) the DNA sequence where the MobM-mediated nick takes place (the oriT(pMV158)); and (iii) the function(s) provided by auxiliary plasmids. MobM belongs to the Pre/Mob family of plasmid-encoded DNA-relaxing proteins (relaxases). Purified MobM protein has been used to assay cleavage conditions on plasmid supercoiled DNA. Some structural features of MobM have been addressed by analytical ultracentrifugation, circular dichroism, thermal denaturation, and fluorescence emission. The protein behaved as a dimer of identical subunits with an ellipsoidal shape. MobM showed a high (about 60%) alpha-helical content and a midpoint denaturation of about 40 degrees C. Cell fractionation assays showed that MobM was associated to the cell membrane. This association was abolished when a great alteration was introduced within a putative coiled-coil located at the C-terminal region of the protein. Emission fluorescence suggested that the three Trp residues of MobM are located within a hydrophobic environment. A molecular model of MobM on the known structure of colicin Ia has been built.
de Antonio
Carmen
C
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Velazquez, 144, E-28006 Madrid, Spain.
Farías
María Eugenia
ME
de Lacoba
Mario García
MG
Espinosa
Manuel
M
D001426:Bacterial Proteins; D002462:Cell Membrane / Q000378:metabolism; D003227:Conjugation, Genetic / Q000235:genetics; D004278:DNA, Superhelical / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D010957:Plasmids / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D021122:Protein Subunits; D021381:Protein Transport; D013291:Streptococcus / Q000737:chemistry; D040681:Structural Homology, Protein
D001426:Bacterial Proteins; D004278:DNA, Superhelical; D021122:Protein Subunits; D004706:Endodeoxyribonucleases; C120220:MobM protein, bacteria