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SRR27151633_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR294/037/SRR29490937/SRR29490937_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR294/031/SRR29490931/SRR29490931_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR271/030/SRR27151630/SRR27151630_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR271/039/SRR27147039/SRR27147039_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR271/036/SRR27147036/SRR27147036_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR271/018/SRR27151418/SRR27151418_1.fastq.gzprimaryOK200GenomicsDuke Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA1050222For over 60 years, salicylic acid (SA) has been known as a plant immune signal required for both basal and systemic acquired resistance (SAR). SA activates these immune responses by reprogramming up to 20% of the transcriptome through the function of NPR1. However, components in the NPR1-signaling hub, which appears as nuclear condensates, and the NPR1-signaling cascade remained elusive due to difficulties in studying transcriptional cofactors whose chromatin associations are often indirect and transient. To overcome this challenge, we applied TurboID to divulge the NPR1-proxiome, which detected almost all known NPR1-interactors as well as new components of transcription-related complexes. Testing of new components showed that chromatin remodeling and histone demethylation contribute to SA-induced resistance. Globally, NPR1-proxiome shares a striking similarity to GBPL3-proxiome involved in SA synthesis, except associated transcription factors (TFs), suggesting that common regulatory modules are recruited to reprogram specific transcriptomes by transcriptional cofactors, like NPR1, through binding to unique TFs. Stepwise greenCUT&RUN analyses showed that, upon SA-induction, NPR1 initiates the transcriptional cascade primarily through association with TGA TFs to induce expression of secondary TFs, predominantly WRKYs. WRKY54 and WRKY70 then play a major role in inducing immune-output genes without interacting with NPR1 at the chromatin. Moreover, a loss of NPR1 condensate formation decreases its chromatin-association and transcriptional activity, indicating the importance of condensates in organizing the NPR1-signaling hub and initiating the transcriptional cascade. This study demonstrates how combinatorial applications of TurboID and stepwise greenCUT&amp RUN transcend traditional genetic methods to globally map signaling hubs and transcriptional cascades.ENAfalseNext-generation mapping of the salicylic acid signaling hub and transcriptional cascade2024-09-242024-09-24PRJNA1050222