- 标题
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Target sequence recognition by a light-activatable bZIP factor, Photozipper
摘要： Photozipper (PZ) is a light-activatable basic leucine zipper (bZIP) protein composed of a bZIP domain and a light-oxygen-voltage-sensing domain of aureochrome-1. Blue light induces dimerization and subsequently increases the affinity of PZ for the target DNA sequence. We prepared site-directed PZ mutants in which Asn131 (N131) in the basic region were substituted with Ala and Gln. N131 mutants showed spectroscopic and dimerization properties almost identical to wild-type PZ, as well as an increase of helical content in the presence of the target sequence. Quantitative analyses by electrophoretic mobility shift assay and quartz crystal microbalance (QCM) measurements demonstrated that the half-maximal effective concentrations of N131 mutants to bind to the target sequence were significantly higher than those of PZ. QCM data also revealed that N131 substitutions accelerated the dissociation speed without affecting the association speed, suggesting that a base-specific interaction of N131 occurred after the association between PZ and DNA. Activation of PZ by illumination decreased both the standard errors and the unstable period of QCM data. Optical control of transcription factors will provide new knowledge of the recognition of the target sequence.
关键词： LOV,DNA-binding,Quartz crystal microbalance (QCM),aureochrome,bZIP
更新于2025-09-23 15:21:01
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Bringing Light to Transcription: The Optogenetics Repertoire
摘要： The ability to manipulate expression of exogenous genes in particular regions of living organisms has profoundly transformed the way we study biomolecular processes involved in both normal development and disease. Unfortunately, most of the classical inducible systems lack fine spatial and temporal accuracy, thereby limiting the study of molecular events that strongly depend on time, duration of activation, or cellular localization. By exploiting genetically engineered photo sensing proteins that respond to specific wavelengths, we can now provide acute control of numerous molecular activities with unprecedented precision. In this review, we present a comprehensive breakdown of all of the current optogenetic systems adapted to regulate gene expression in both unicellular and multicellular organisms. We focus on the advantages and disadvantages of these different tools and discuss current and future challenges in the successful translation to more complex organisms.
关键词： transcription,optogenetics,cryptochrome,phytochrome B,LOV,gene expression,UVR8,light
更新于2025-09-09 09:28:46