Complementation of an Escherichia coli DnaK defect by Hsc70-DnaK chimeric proteins.
Escherichia coli DnaK and rat Hsc70 are members of the highly conserved 70-kDa heat shock protein (Hsp70) family that show strong sequence and structure similarities and comparable functional properties in terms of interactions with peptides and unfolded proteins and cooperation with cochaperones. We show here that, while the DnaK protein is, as expected, able to complement an E. coli dnaK mutant strain for growth at high temperatures and lambda phage propagation, Hsc70 protein is not. However, an Hsc70 in which the peptide-binding domain has been replaced by that of DnaK is able to complement this strain for both phenotypes, suggesting that the peptide-binding domain of DnaK is essential to fulfill the specific functions of this protein necessary for growth at high temperatures and for lambda phage replication. The implications of these findings on the functional specificities of the Hsp70s and the role of protein-protein interactions in the DnaK chaperone system are discussed.
Suppini
Jean-Philippe
JP
University P. & M. Curie, CNRS, 96 Bd. Raspail, 75006 Paris, France. ladjimi@ccr.jussieu.fr
Amor
Mouna
M
Alix
Jean-Hervé
JH
Ladjimi
Moncef M
MM
D000818:Animals; D010582:Bacteriophage lambda / Q000254:growth & development; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; C063842:dnaK protein, E coli