Specific in vivo protein-protein interactions between Escherichia coli SOS mutagenesis proteins.
One of the components of the RecA-LexA-controlled SOS response in Escherichia coli cells is an inducible error-prone DNA replication pathway that results in a substantial increase in the mutation rate. It is believed that error-prone DNA synthesis is performed by a multiprotein complex that is formed by UmuC, UmuD', RecA, and probably DNA polymerase III holoenzyme. It is postulated that the formation of such a complex requires specific interactions between these proteins. We have analyzed the specific protein-protein interactions between UmuC, UmuD, and UmuD' fusion proteins, using a Saccharomyces cerevisiae two-hybrid system. In agreement with previous in vitro data, we have shown that UmuD and UmuD' are able to form both homodimers (UmuD-UmuD and UmuD'-UmuD') and a heterodimer (UmuD-UmuD'). Our data show that UmuC fusion protein is capable of interacting exclusively with UmuD' and not with UmuD. Thus, posttranslational processing of UmuD into UmuD' is a critical step in SOS mutagenesis, enabling only the latter protein to interact with UmuC. Our data seem to indicate that the integrity of the entire UmuC sequence is essential for UmuC-UmuD' heterotypic interaction. Finally, in our studies, we used three different UmuC mutant proteins: UmuC25, UmuC36, and UmuC104. We have found that UmuC25 and UmuC36 are not capable of associating with UmuD'. In contrast, UmuC104 protein interacts with UmuD' protein with an efficiency identical to that of the wild-type protein. We postulate that UmuC104 protein might be defective in interaction with another, unknown protein essential for the SOS mutagenesis pathway.
Jonczyk
P
P
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
Nowicka
A
A
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013014:SOS Response, Genetics / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; C078202:UmuC protein, E coli; D004259:DNA-Directed DNA Polymerase; C055218:UmuD protein, E coli; D001616:beta-Galactosidase