JASPer controls interphase histone H3S10 phosphorylation by chromosomal kinase JIL-1 in Drosophila

In flies, the chromosomal kinase JIL-1 is responsible for most interphase histone H3S10 phosphorylation and has been proposed to protect active chromatin from acquiring heterochromatic marks, such as dimethylated histone H3K9 (H3K9me2) and HP1. Here, we show that JIL-1’s targeting to chromatin depends on a PWWP domain-containing protein JASPer (JIL-1 Anchoring and Stabilizing Protein). JASPer-JIL-1 (JJ)-complex is the major form of kinase in vivo and is targeted to active genes and telomeric transposons via binding of the PWWP domain of JASPer to H3K36me3 nucleosomes, to modulate transcriptional output. JIL-1 and JJ-complex depletion in cycling cells lead to small changes in H3K9me2 distribution at active genes and telomeric transposons. Finally, we identify interactors of the endogenous JJ-complex and propose that JIL-1 not only prevents heterochromatin formation but also coordinates chromatin-based regulation in the transcribed part of the genome.


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Sample size
Sample sizes were not determined by statistical test. Generally, => 3 biological replicates were used for ChIP-seq and => 4 for RNA-seq, based on community standards.
Data exclusions RNA-seq samples with low number of reads were excluded.

Replication
All ChIP replicates for each antibody have been prepared on independent Chromatin preparations, indicated by the number of replicates (n). When possible different antibodies against one antigen (JIL-1, JASPer) have been used (in general 3 independent replicates). The libraries of biological replicates have been prepared and sequenced together or separately. The RNAi experiments for ChIP-seq analysis have been done with 2 independent dsRNA constructs for each target knock down as described in the methods.

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