CDC6 is an essential gene of yeast Saccharomyces cerevisiae. It has a pivotal function in the initiation of DNA replication by interacting with ORC complex and loading MCM proteins to ARS sequences. ...Besides in S phase of the cell cycle, Cdc6 protein has a role in the suppression of nuclear division. Undestanding function of proteins interacting with Cdc6 may tell us a lot about the biochemical pathways in the eukaryotic cell as well as about the protein itself. Since Cdc6 is an unstable and difficult to be monitored with conventional biochemical methdods, we chose the two-hybrid system. Exploting the phenomena that transcription is induced by two distinct domains, this system provides a toll for monitoring physical interactions of Cdc6 with otherproteins in the cell. Four gene products interacting with Cdc6 were detectedČ YOR279, ECM11, YNL279 and Ty1A element. ECM11 codes for the nonessential protein involved in the cell wall synthesis, YNL201 codes for thenonessential protein, probably involved in the carbon metabolism, YOR279 codes for protein of completely unknown function with no significant similarity with any known protein. We found that ćknock outć of this protein is viable. The strenth of interation with Cdc6 bait was measured as beta-galactosidase activity. The strongest interaction was observed with ECM11, YNL201 and YOR279 interacts slightly weaker. The weakest beta-galactosidase activity was obtained by Ty1A element. The genetic interactions with CDC6 were detected by complementation tests with cdc-1 termosensitive strain. The strongest suppression of cdc-1 mutation was observed by Ty1A element, the slight one with ECM11 and YNL201 but no suppression of thermosensitive mutation was detected for YOR279. For new gene products interacting with Cdc6 were found by the two-hybrid method and none ofthem was described in the literature before. (Abstract truncated at 2000 characters).
Type of material - master's thesis
Publication and manufacture - Ljubljana : [A. Bedina Zavec], 1999
magistrske naloge BEDINA Zavec A. Interakcije IN: 099901041
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