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Abstract：
Heat-shock protein beta1 (HSPB1) is a member of the small HSP family, downregulated in osteoarthritis (OA) chondrocytes and demonstrated the capacity to serve as an RNA-binding protein (RBP). This work aimed to explore the profile of HSPB1 bound RNA and reveal the potential regulation mechanism of HSPB1 in OA. In this work, we captured an unbiased HSPB1-RNA interaction map in Hela cells using the iRIP-seq. The results demonstrated that HSPB1 interacted with plentiful of mRNAs and genomic location toward the CDS region. Functional enrichment of HSPB1-related peaks showed the involvement in gene expression, translation initiation, cellular protein metabolic process, and nonsense-mediated decay. HOMER software analysis showed that HSPB1 bound peaks were over-represented in GAGGAG sequences. In addition, ABLIRC and CIMS algorithm indicated that HSPB1 bound to AU-rich motifs and the proportion of AU-rich peaks in 3\' UTR were slightly higher than that in other regions. Moreover, HSPB1-binding targets analysis revealed several gens were associated with OA including EGFR, PLEC, COL5A1, and ROR2. The association of OA-related mRNAs to HSPB1 was additionally confirmed in OA tissues by the quantitative RIP-PCR experiments. Further experiment demonstrated the downregulation of HSPB1 in OA tissues. In conclusion, our current study confirmed HSPB1 as an RNA-binding protein and revealed its potential function in the pathological process of OA, providing a reliable insight to further investigate the molecular regulation mechanism of HSPB1 in OA.
摘要：
热休克蛋白β1（HSPB1）是小的HSP家族的成员，在骨关节炎（OA）软骨细胞中下调，并证明了作为RNA结合蛋白（RBP）的能力。本工作旨在探索HSPB1结合RNA的谱图，揭示HSPB1在OA中的潜在调控机制。在这项工作中,我们使用iRIP-seq在Hela细胞中捕获了无偏倚的HSPB1-RNA相互作用图谱。结果表明，HSPB1与大量的mRNAs相互作用，基因组位于CDS区。HSPB1相关峰的功能富集显示参与基因表达，翻译启动,细胞蛋白质代谢过程，和无意义介导的衰变。HOMER软件分析显示HSPB1结合峰在GAGGAG序列中过度表示。此外,ABLIRC和CIMS算法表明，HSPB1与富含AU的基序结合，3'UTR中富含AU的峰比例略高于其他区域。此外，HSPB1结合靶标分析显示几个基因与OA相关，包括EGFR，PLEC,COL5A1和ROR2。通过定量RIP-PCR实验在OA组织中进一步证实了OA相关mRNA与HSPB1的关联。进一步实验证明了OA组织中HSPB1的下调。总之,我们目前的研究证实HSPB1是一种RNA结合蛋白，并揭示了其在OA病理过程中的潜在功能，为进一步研究HSPB1在OA中的分子调控机制提供了可靠的见解。
