Study On The Regulation Of C1orf109 280 In Cellular Stress

C1orf109 is a novel gene has four transcripts,which structure and function are remained unknown.Previous study demonstrated that the protein encoded by C1orf109 280 is located in the nucleus,and under UV radiation localization of C1orf109280 transcript can be detected in the cytoplasm.It might play an important role in tumor cells under cellular stress response.It was found that the absence of external stimulation,C1orf109 280 transcript is mainly located in the nucleus.After UV radiation,the results showed that the transcript is located in the cytoplasm which suggested that it may be occurred under cellular stress response.Therefore,in this report,we used localization analysis to explore the function of C1 orf 109 280 transcript and discuss its regulatory mechanism in the process of trans-localization from nucleus to cytoplasm in the cellular stress.In order to further investigate,we explored the function and mechanism of C1orf109 280 transcripts in tumor cells under the stress response.Firstly,we treated cells with high concentration of hydrogen peroxide for short time(500 mol/L,2h)and performed immunofluorescence,real-time PCR,and Western-blot technique to identified the localization of C1orf109 280 transcript,the results demonstrated that the localization C1orf109 280 is transferred from the nucleus to the cytoplasm.Secondly,we determined the sub-cellular localization of C1orf109 280 transcripts in mitochondria by using mitochondria separation technology,immunofluorescence and Western-blot technique under the cellular stress,and the protein expression level was not altered.On the basis of obtaining the C1orf109 280 transcriptional we constructed a stable cell line,and the cell viability was detected by CCK-8.The results revealed that viability of stable cell lines was significantly reduced.Under cellular stress,the results revealed that the activity/viability of over-expressed C1orf109 280 transcriptional cells was significantly lower as compared with the control group,furthermore,by using the live cell workstation technique and under TNF-? stress conditions we found that C1orf109 280 transcript cells showed follicular ruptured and the death pattern was significantly different as compared with the control group and morphology of death was remained similar as in necroptosis.Finally,we conducted silver staining technique,mass spectrometry sequencing,Western-blot technique and co-immunoprecipitation to clarify theinteraction between C1orf109 280 transcriptional transcript and phosphate mutant enzyme 5(PGAM5).In the programmed cell necroptosis signal pathway,the RIP3-MLKL pathway is clearly studied.PGAM5 is a downstream molecule regulated by this pathway,and is the key protein to determine programmed cell necroptosis.In summary,C1orf109 280 is normally located in the nucleus,but under stress conditions,C1orf109 280 trans-located in the cytoplasm and localized in the mitochondria,which affected cellular viability.And TNF-? induction can cause cell swelling/ expansion,follicle rupture and death,and interact with PGAM5.Therefore,we suggested that the transcript of C1orf109 280 has a certain effect on cell death under the cellular stress.