Interaction trap experiment with CDC6

Abstract

CDC6 is an essential gene of yeast Saccharomyces cerevisiae. Although DNA sequence of the gene is available for a long time, biochemical function of Cdc6 protein in the cell cycle remains unclear. Using the interaction trap experiment we were looking for proteins interacting specifically with Cdc6. Four gene products interacting with Cdc6 were detected. By sequence analysis we found that ECM11 codes for the protein involved in the cell wall synthesis, YNL201 codes for the protein of unknown function, probably involved in the carbon metabolism, YOR279 codes for protein of completely unknown function with no significant similarity with any known protein, and the interaction with Ty1 retrotransposition element was also found. The strongest interaction with Cdc6 bait measured as beta-galactosidase activity was observed with ECM11 and YNL201; YOR279 interacts slightly weaker. The weakest beta-galactosidase activity was obtained by Ty1A element. The strongest suppression of cdc6-1 mutation was observed by Ty1A element, the slight one with ECM11 and YNL201 but no suppression of thermosensitive mutation was detected for YOR279.

Intrinsically Disoredered Proteins in Replication Process

“…Genetic interactions between Cdc6 and Ecm11 were also observed. Moderate supression of cdc6-1 mutation by overexpression of ECM11 was detected [72] and deletion of ECM11 in cdc6-1 genetic background enhances thermo-sensitivity of cdc6-1 mutation (Zavec AB, unpublished result). These data suggest direct involving of Ecm11 in initiation of DNA replication process.…”

“…In the two-hybrid screen it was found out that Ecm11 strongly interacts with Cdc6 that has a pivotal role in the initiation of DNA replication [72]. Genetic interactions between Cdc6 and Ecm11 were also observed.…”