DNA Methylation of a GC Repressor Element in the Smooth Muscle Myosin Heavy Chain Promoter Facilitates Binding of the Notch-Associated Transcription Factor, RBPJ/CSL1

OBJECTIVE: The goal of the present study was to identify novel mechanisms that regulate smooth muscle cell (SMC) differentiation marker gene expression. APPROACH AND RESULTS: We demonstrate that the CArG-containing regions of many SMC-specific promoters are imbedded within CpG islands. A previously identified GC repressor element in the SM myosin heavy chain (MHC) promoter was highly methylated in cultured aortic SMC but not in the aorta, and this difference was inversely correlated with SM MHC expression. Using an affinity chromatography/mass spectroscopy-based approach, we identified the multifunctional Notch transcription factor, recombination signal binding protein for immunoglobulin κ J region (RBPJ), as a methylated GC repressor-binding protein. RBPJ protein levels and binding to the endogenous SM MHC GC repressor were enhanced by platelet-derived growth factor-BB treatment. A methylation mimetic mutation to the GC repressor that facilitated RBPJ binding inhibited SM MHC promoter activity as did overexpression of RBPJ. Consistent with this, knockdown of RBPJ in phenotypically modulated human aortic SMC enhanced endogenous SMC marker gene expression, an effect likely mediated by increased recruitment of serum response factor and Pol II to the SMC-specific promoters. In contrast, the depletion of RBPJ in differentiated transforming growth factor-β-treated SMC inhibited SMC-specific gene activation, supporting the idea that the effects of RBPJ/Notch signaling are context dependent. CONCLUSIONS: Our results indicate that methylation-dependent binding of RBPJ to a GC repressor element can negatively regulate SM MHC promoter activity and that RBPJ can inhibit SMC marker gene expression in phenotypically modulated SMC. These results will have important implications on the regulation of SMC phenotype and on Notch-dependent transcription

RF IC performance optimization by synthesizing optimum inductors

Even with optimal system design and careful choice of topology for a particular RF application, large amounts of energy are often wasted due to low-quality passives, especially inductors. Inductors have traditionally been difficult to integrate due to their inherent low quality factors and modelling complexity. Furthermore, although many different inductor configurations are available for an RF designer to explore, support for integrated inductors in electronic design automation tools and process design kits has been very limited in the past. In this chapter, a recent advance in technology-aware integrated inductor design is presented, where drawbacks of the integrated inductor design are addressed by introducing an equation-based inductor synthesis algorithm. The intelligent computation technique aims to allow RF designers to optimize integrated inductors, given the inductor center frequency dictated by the device application, and geometry constraints. This does not only lay down a foundation for system-level RF circuit performance optimization, but, because inductors are often the largest parts of an RF system, it also allows for optimal usage of chip real estate