裂解和聚腺苷酸化特异性因子(CPSF)是一种蛋白质复合物,在mRNA3'端形成中起着重要的生化作用。包括poly(A)信号识别和poly(A)位点的切割。然而,在多细胞真核生物中,其在生物体水平上的生物学功能大多是未知的。植物CPSF73的研究受到拟南芥(拟南芥)纯合突变体AtCPSF73-I和AtCPSF73-II的致死性的阻碍。这里,我们使用poly(A)标签测序(PAT-seq)研究了AtCPSF73-I和AtCPSF73-II在AN3661治疗的拟南芥中的作用,AN3661是一种对与植物CPSF73同源的寄生虫CPSF73具有特异性的抗疟药.在AN3661处理的培养基上直接种子萌发是致命的;然而,用AN3661处理的7天龄幼苗存活。AN3661针对AtCPSF73-I和AtCPSF73-II,通过协调基因表达和poly(A)位点选择来抑制生长。功能富集分析表明,乙烯和生长素的积累共同抑制了初生根的生长。AN3661受影响的poly(A)信号识别,导致较低的U型富信号使用率,导致转录连读,并增加了远端poly(A)位点的使用。在3'UTR延长的转录物中发现了许多microRNA靶标;这些miRNA可以通过切割间接调节这些靶标的表达。总的来说,这项工作表明,AtCPSF73在共转录调控中起着重要的作用,影响拟南芥的生长和发育。
Cleavage and polyadenylation specificity factor (CPSF) is a protein complex that plays an essential biochemical role in mRNA 3\'-end formation, including poly(A) signal recognition and cleavage at the poly(A) site. However, its biological functions at the organismal level are mostly unknown in multicellular eukaryotes. The study of plant CPSF73 has been hampered by the lethality of Arabidopsis (Arabidopsis thaliana) homozygous mutants of AtCPSF73-I and AtCPSF73-II. Here, we used poly(A) tag sequencing to investigate the roles of AtCPSF73-I and AtCPSF73-II in Arabidopsis treated with AN3661, an antimalarial drug with specificity for parasite CPSF73 that is homologous to plant CPSF73. Direct seed germination on an AN3661-containing medium was lethal; however, 7-d-old seedlings treated with AN3661 survived. AN3661 targeted AtCPSF73-I and AtCPSF73-II, inhibiting growth through coordinating gene expression and poly(A) site choice. Functional enrichment analysis revealed that the accumulation of ethylene and auxin jointly inhibited primary root growth. AN3661 affected poly(A) signal recognition, resulted in lower U-rich signal usage, caused transcriptional readthrough, and increased the distal poly(A) site usage. Many microRNA targets were found in the 3\' untranslated region lengthened transcripts; these miRNAs may indirectly regulate the expression of these targets. Overall, this work demonstrates that AtCPSF73 plays important part in co-transcriptional regulation, affecting growth, and development in Arabidopsis.