Aday saat düzenleyici gen olarak ınterferon ile indüklenmiş transmembran protein 1

Daily rhythms, known as the circadian rhythm, are ever-present in the living world, driving the sleep–wake cycle and many other physiological changes. Most of the physiological pathways are controlled by the interaction between different signal transduction pathway and core clock mechanism. However, many aspects of circadian molecular physiology remain unexplained. In the last two decades, several groups have identified "clock genes" that interact to generate underlying molecular oscillations and, in turn, regulate the different physiological variables. At the molecular level the CLOCK/BMAL1 transcriptional complex lies at the core of the molecular clock. These proteins bind E-box elements in the promoters of target genes. The Period and Cryptochrome gene families are prominent among these targets, and their products ultimately repress CLOCK/BMAL1 activity and their own transcription. A second loop regulates Bmal1 expression through the opposing actions of the REV–ERB and ROR nuclear receptor protein families. A previous study used a simple "machine learning" approach to identify new clock genes by searching the genome for candidate genes that share clock-like features such as cycling, broad-based tissue RNA expression, in vitro circadian activity, genetic interactions, and homology across species. This study resulted in identification of a gene, Interferon Induced Transmembrane Protein 1 (IFITM1), may also act as in a clock function. In this thesis work, I showed that IFITM1 protein specially interacts with CRY1 proteins using mammalian two-hybrid and co-immunoprecipitation. Further studies showed last 20 amino acids in CRY1 C-termini are responsible for the interaction with IFITM1. To evaluate the functional consequences of these physical interactions, I monitored Per1::luciferase activity in unsynchronized HEK 293T cells transiently transfected with Clock/Bmal1, Cry1 and Ifitm1. It is known that Per1::luc reporter activity is enhanced by CLOCK/BMAL1 transfection but repressed by the overexpression of Cry1. When all of the constructs were transfected in HEK 293T cells, we observed Ifitm1 inhibits repressor activity of the CRY1. In this study for the first time, I provided evidence that IFTIM1 interacts with CRY1 and such interaction v inhibits the activity of CRY1. This study improves our understanding on how clock works and will enable us to develop new drugs curing clock-related diseases such as insomnia, diabetes, obesity, jetlag and psychological disorders.