role of architecture in the function and specificity of two notch-regulated transcriptional enhancer modules角色的建筑功能和特异性两notch-regulated转录增强器模块.pdfVIP

Role of Architecture in the Function and Specificity of Two Notch-Regulated Transcriptional Enhancer Modules ¤ Feng Liu , James W. Posakony* Cell and Developmental Biology Section, Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America Abstract In Drosophila melanogaster, cis-regulatory modules that are activated by the Notch cell–cell signaling pathway all contain two types of transcription factor binding sites: those for the pathway’s transducing factor Suppressor of Hairless [Su(H)] and those for one or more tissue- or cell type–specific factors called ‘‘local activators.’’ The use of different ‘‘Su(H) plus local activator’’ motif combinations, or codes, is critical to ensure that only the correct subset of the broadly utilized Notch pathway’s target genes are activated in each developmental context. However, much less is known about the role of enhancer ‘‘architecture’’—the number, order, spacing, and orientation of its component transcription factor binding motifs—in determining the module’s specificity. Here we investigate the relationship between architecture and function for two Notch-regulated enhancers with spatially distinct activities, each of which includes five high-affinity Su(H) sites. We find that the first, which is active specifically in the socket cells of external sensory organs, is largely resistant to perturbations of its architecture. By contrast, the second enhancer, active in the ‘‘non-SOP’’ cells of the proneural clusters from which neural precursors arise, is sensitive to even simple rearrangements of its transcription factor binding sites, responding with both loss of normal specificity and striking ectopic activity. Thus, diverse cryptic specificities can be inherent in an enhancer’s partic