Research On The C1orf109 Binding With Cyclin D1 Promoter And The Sequence Signature

In previous research, our research group had found a new protein called c1orf109. Previous results showed that c1orf109 has three phosphorylation site and is the substrate of CK kinase. And the fact that the protein having high expression level in multiple cancer cell lines indicates there is potential relationship between c1orf109 and the canceration process. Results also indicated that c1orf109 show affinity with cyclin D1 promoter.To stimulate if this affinity is the way that c1orf109 regulate the expression of cyclin D1 and explain the high level of c1orf109 in cancer cells. This study mainly research if there is interaction between c1orf109 and cyclin D1’s promoter sequences and could regulate cyclin D1 protein expression, and the main interac region on cyclin D1’s promoter.The study first use bioinformatics methods to analyze the promoter region of cyclin D1 and compared with regions which have combination with c1orf109 protein, and get six regions of the promoter. Then these cyclin D1’s promoter region were combined the with luciferase report plasmid PGL3-basic, and express different levels of luciferase in He La under the stimulation of endogenic c1orf109 protein. To create different levels of c1orf109, the study used RNA interference and western blot results showed that all three of si RNA could knockdown c1orf109. Finally, luciferase report plasmids combined with cyclin D1’s promoter were transfect to He La cells which had different c1orf109 level by RNA interference and tested the luciferase level. The results showed that luciferase report plasmids recombined with-881-15 bp and-316-15 bp cyclin D1’s promoter express high level of luciferase, and show down-regulation when the level of c1orf109 get down.Those results proved the fact that c1orf109 protein could make cyclin D1’s expression upregulated, and this regulation is achieved by the interaction between c1orf109 protein and the promoter of cyclin D1. Experiments finally verified the region that c1orf109 protein could effect on cyclin D1 promoter, these results could provide clue for further research on the mechanism of the regulation and the sequence of regulation element which is affected by c1orf109 protein.