DmsD, a Tat system specific chaperone, interacts with other general chaperones and proteins involved in the molybdenum cofactor biosynthesis.
Many bacterial oxidoreductases depend on the Tat translocase for correct cell localization. Substrates for the Tat translocase possess twin-arginine leaders. System specific chaperones or redox enzyme maturation proteins (REMPs) are a group of proteins implicated in oxidoreductase maturation. DmsD is a REMP discovered in Escherichia coli, which interacts with the twin-arginine leader sequence of DmsA, the catalytic subunit of DMSO reductase. In this study, we identified several potential interacting partners of DmsD by using several in vitro protein-protein interaction screening approaches, including affinity chromatography, co-precipitation, and cross-linking. Candidate hits from these in vitro findings were analyzed by in vivo methods of bacterial two-hybrid (BACTH) and bimolecular fluorescence complementation (BiFC). From these data, DmsD was confirmed to interact with the general molecular chaperones DnaK, DnaJ, GrpE, GroEL, Tig and Ef-Tu. In addition, DmsD was also found to interact with proteins involved in the molybdenum cofactor biosynthesis pathway. Our data suggests that DmsD may play a role as a "node" in escorting its substrate through a cascade of chaperone assisted protein-folding maturation events.
Li
Haiming
H
Department of Biological Sciences, Faculty of Science, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Chang
Limei
L
Howell
Jenika M
JM
Turner
Raymond J
RJ
D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003067:Coenzymes / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008667:Metalloproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D011621:Pteridines / Q000737:chemistry
D002352:Carrier Proteins; D003067:Coenzymes; D029968:Escherichia coli Proteins; D047908:Intracellular Signaling Peptides and Proteins; D007506:Iron-Sulfur Proteins; D026901:Membrane Transport Proteins; D008667:Metalloproteins; D018832:Molecular Chaperones; D000090006:Molybdenum Cofactors; D010088:Oxidoreductases; D011621:Pteridines; C434268:DmsD protein, E coli; C408585:twin-arginine translocase complex, E coli; C022261:molybdopterin; C045664:dimethyl sulfoxide reductase