Position-specific binding of FUS to nascent RNA regulates mRNA length

• Published in: Genes & development Vol. 29; no. 10; pp. 1045 - 1057
• Main Authors: Masuda, Akio, Takeda, Jun-ichi, Okuno, Tatsuya, Okamoto, Takaaki, Ohkawara, Bisei, Ito, Mikako, Ishigaki, Shinsuke, Sobue, Gen, Ohno, Kinji
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 15.05.2015
• Subjects: Animals; Cell Line, Tumor; Cleavage And Polyadenylation Specificity Factor - metabolism; Gene Expression Regulation; Gene Knockdown Techniques; HEK293 Cells; Humans; Mice; Polyadenylation; Protein Binding; Research Paper; RNA - metabolism; RNA Polymerase II - metabolism; RNA, Messenger - metabolism; RNA-Binding Protein FUS - metabolism; Transcriptome; mRNA length; FUS; RNA polymerase II; CLIP; alternative polyadenylation
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.255737.114

More than half of all human genes produce prematurely terminated polyadenylated short mRNAs. However, the underlying mechanisms remain largely elusive. CLIP-seq (cross-linking immunoprecipitation [CLIP] combined with deep sequencing) of FUS (fused in sarcoma) in neuronal cells showed that FUS is frequently clustered around an alternative polyadenylation (APA) site of nascent RNA. ChIP-seq (chromatin immunoprecipitation [ChIP] combined with deep sequencing) of RNA polymerase II (RNAP II) demonstrated that FUS stalls RNAP II and prematurely terminates transcription. When an APA site is located upstream of an FUS cluster, FUS enhances polyadenylation by recruiting CPSF160 and up-regulates the alternative short transcript. In contrast, when an APA site is located downstream from an FUS cluster, polyadenylation is not activated, and the RNAP II-suppressing effect of FUS leads to down-regulation of the alternative short transcript. CAGE-seq (cap analysis of gene expression [CAGE] combined with deep sequencing) and PolyA-seq (a strand-specific and quantitative method for high-throughput sequencing of 3' ends of polyadenylated transcripts) revealed that position-specific regulation of mRNA lengths by FUS is operational in two-thirds of transcripts in neuronal cells, with enrichment in genes involved in synaptic activities.