H3-ChIP-Seq and RNA-Seq in A.thaliana SPT16 phospho-mutant lines

FACT consisting of SSRP1 and SPT16 is conserved in a wide variety of plant species. FACT can disassemble and reassemble nucleosomes; thereby it is involved in both overcoming and maintaining the nucleosomal barrier to DNA-dependent processes in the chromatin context. There is only limited information about the potential regulation of FACT function via post-translational modifications. We mutated several identified SPT16 phosphorylation sites followed by RNA-Seq and H3-ChIP-Seq as a measurement for total genome-wide nucleosome occupany in the mutant lines. The expression of a non-phosphorylatable SPT16 variant results in a distinct rearrangement of H3.由SSRP1与SPT16构成的FACT（Facilitates Chromatin Transcription）在诸多植物物种中均高度保守。FACT能够介导核小体的解离与重建，进而参与染色质微环境中DNA依赖型生物学过程所涉及的核小体屏障的克服与维持。目前，关于通过翻译后修饰调控FACT功能的相关研究信息仍较为有限。我们对已鉴定出的数个SPT16磷酸化位点进行突变，并借助RNA测序（RNA-Seq）与H3染色质免疫沉淀测序（H3-ChIP-Seq）检测突变株系的全基因组范围核小体占据率。表达非磷酸化型SPT16变体可引发H3组蛋白的显著重排。