Therefore, it seems likely that each 5 UTR hardly influenced the corresponding promoter activity of the CRELD2 and ALG12 promoter constructs in our assay system.CRELD2 and ALG12 genes possess 5 UTR and 3 UTR respectively though their effects on transcription are not elucidated yet. Further characterization of these regions would reveal regula tions of CRELD2 and ALG12 nearly mRNA expression. Using various deletion mutation constructs, we showed that three suppressive sites in the CRELD2 ALG12 gene pair play a crucial role in interfering with Tg responsiveness. Interestingly, the deletion of all three of these suppressive sites was required in order to restore the responsiveness to Tg. These results imply that these suppressive sites are not only important in maintaining basal promoter activity, but that they synergistically counteract the ERSE mediated transcriptional activity.

Among these sites, the most proximal to the ALG12 promoter contains a conserved response Inhibitors,Modulators,Libraries element that Ets family transcriptional factors recognize. Inhibitors,Modulators,Libraries Ets transcription factors consist of approximately 30 family members and share a highly conserved DNA binding domain. It has been reported that these factors are involved in regulat GSK-3 ing a variety of biological processes including develop ment, differentiation and inflammation. In the site II, there are putative YY1 and MAZ binding sites judged from some databases such as SwissRegulon, but the precise roles remain to be determined. On the contrary, we are unable to find any unique sequences in the sites I.

Further studies characterizing each of these suppres sive sites are required in order to understand the complex transcriptional regulation of the CRELD2 ALG12 gene pair. Jones PL et al. reported that murine manganese superoxide dismutase gene is regu Inhibitors,Modulators,Libraries lated through a complex intronic enhancer involving C EBP b and NF B. Donati G et al. demonstrated that ER stress triggers dynamic modification of chroma tin components and transcriptional factors under ER stress. Therefore, we should focus on other aspects such as local chromatin remodeling and histone modifi cations within the CRELD2 and ALG12 genes in addition to the 5 flanking sequences in this inter genic region. Furthermore, other approaches should be employed to elucidate the discrepancy between the expression Inhibitors,Modulators,Libraries levels of both intrinsic mRNAs and the pro moter activities of their full intergenic region under ER stress conditions.

Among the bidirectional gene sellectchem pairs characterized in mammalian cells, Surf1 Surf2, Reql4 Lrrc14, PDCD10 SERPINI1 and Thox DUOXA gene pairs seem to share their intergenic region equally because mutations in the transcription factor binding sites decline those promoter activities equally. In con trast, the transcriptional regulations of C2ORF34 PREPL, Sarsm Mrps12 and HAND2 DEIN are asym metric.