Comparative Oncology Laboratory, Schools of Veterinary Medicine and Medicine, University of California, Davis, California, USA.
Comparative Oncology Laboratory, Schools of Veterinary Medicine and Medicine, University of California, Davis, California, USA.
J Invest Dermatol. 2020 Nov;140(11):2166-2177.e6. doi: 10.1016/j.jid.2020.03.942. Epub 2020 Mar 31.
p63 is expressed from two promoters and produces two N-terminal isoforms, TAp63 and ΔNp63. Alternative splicing creates three C-terminal isoforms p63α, p63β, and p63δ, whereas alternative polyadenylation (APA) in coding sequence creates two more C-terminal isoforms p63γ and p63ε. Although several transcription factors have been identified to differentially regulate the N-terminal p63 isoforms, it is unclear how the C-terminal p63 isoforms are regulated. Thus, we determined whether PABPN1, a key regulator of APA, may differentially regulate the C-terminal p63 isoforms. We found that PABPN1 deficiency increases p63γ mRNA through APA in coding sequence. We also found that PABPN1 is necessary for p63α translation by modulating the binding of translation initiation factors eIF4E and eIF4G to p63α mRNA. Moreover, we found that the p53 family, especially p63α, regulates PABPN1 transcription, suggesting that the mutual regulation between p63 and PABPN1 forms a feedback loop. Furthermore, we found that PABPN1 deficiency inhibits keratinocyte cell growth, which can be rescued by ectopic ΔNp63α. Finally, we found that PABPN1 controls the terminal differentiation of HaCaT keratinocytes by modulating ΔNp63α expression. Taken together, our findings suggest that PABPN1 is a key regulator of the C-terminal p63 isoforms through APA in coding sequence and mRNA translation and that the p63-PABPN1 loop modulates p63 activity and the APA landscape.
p63 由两个启动子表达,并产生两个 N 端同工型,TAp63 和 ΔNp63。选择性剪接产生三个 C 端同工型 p63α、p63β 和 p63δ,而编码序列中的选择性多聚腺苷酸化(APA)则产生另外两个 C 端同工型 p63γ 和 p63ε。虽然已经鉴定出几种转录因子来差异调节 N 端 p63 同工型,但尚不清楚 C 端 p63 同工型是如何被调节的。因此,我们确定了 APA 的关键调节因子 PABPN1 是否可能差异调节 C 端 p63 同工型。我们发现 PABPN1 缺乏通过编码序列中的 APA 增加 p63γ mRNA。我们还发现 PABPN1 通过调节翻译起始因子 eIF4E 和 eIF4G 与 p63α mRNA 的结合来促进 p63α 翻译是必需的。此外,我们发现 p53 家族,特别是 p63α,调节 PABPN1 的转录,这表明 p63 和 PABPN1 之间的相互调节形成了一个反馈回路。此外,我们发现 PABPN1 缺乏抑制角质形成细胞的生长,这可以通过异位表达 ΔNp63α 得到挽救。最后,我们发现 PABPN1 通过调节 ΔNp63α 的表达来控制 HaCaT 角质形成细胞的终末分化。总之,我们的研究结果表明,PABPN1 通过编码序列中的 APA 和 mRNA 翻译作为 C 端 p63 同工型的关键调节因子,并且 p63-PABPN1 环调节 p63 活性和 APA 景观。