Competitive regulation of alternative splicing and alternative polyadenylation by hnRNP H and CstF64 determines acetylcholinesterase isoforms.

Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE

Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.

Alternative Splicing* ; Polyadenylation*; Acetylcholinesterase/*genetics ; Acetylcholinesterase/*metabolism ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H/*genetics ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H/*metabolism ; RNA-Binding Proteins/*genetics ; RNA-Binding Proteins/*metabolism; Base Sequence ; Binding, Competitive ; Caco-2 Cells ; Cell Line ; Cleavage Stimulation Factor ; Exons ; GPI-Linked Proteins/genetics ; GPI-Linked Proteins/metabolism ; Gene Expression Regulation, Enzymologic ; Gene Knockdown Techniques ; HEK293 Cells ; HeLa Cells ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H/antagonists & inhibitors ; Humans ; Isoenzymes/genetics ; Isoenzymes/metabolism ; Models, Biological ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Regulatory Elements, Transcriptional

Acetylcholinesterase (AChE), encoded by the ACHE gene, hydrolyzes the neurotransmitter acetylcholine to terminate synaptic transmission. Alternative splicing close to the 3΄ end generates three distinct isoforms of AChET, AChEH and AChER. We found that hnRNP H binds to two specific G-runs in exon 5a of human ACHE and activates the distal alternative 3΄ splice site (ss) between exons 5a and 5b to generate AChET. Specific effect of hnRNP H was corroborated by siRNA-mediated knockdown and artificial tethering of hnRNP H. Furthermore, hnRNP H competes for binding of CstF64 to the overlapping binding sites in exon 5a, and suppresses the selection of a cryptic polyadenylation site (PAS), which additionally ensures transcription of the distal 3΄ ss required for the generation of AChET. Expression levels of hnRNP H were positively correlated with the proportions of the AChET isoform in three different cell lines. HnRNP H thus critically generates AChET by enhancing the distal 3΄ ss and by suppressing the cryptic PAS. Global analysis of CLIP-seq and RNA-seq also revealed that hnRNP H competitively regulates alternative 3΄ ss and alternative PAS in other genes. We propose that hnRNP H is an essential factor that competitively regulates alternative splicing and alternative polyadenylation.

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0 (CSTF2T protein, human)
0 (Cleavage Stimulation Factor)
0 (GPI-Linked Proteins)
0 (Heterogeneous-Nuclear Ribonucleoprotein Group F-H)
0 (Isoenzymes)
0 (RNA, Messenger)
0 (RNA-Binding Proteins)
EC 3.1.1.7 (ACHE protein, human)
EC 3.1.1.7 (Acetylcholinesterase)

Date Created: 20170210 Date Completed: 20171030 Latest Revision: 20201209

20231215

PMC5388418

10.1093/nar/gkw823

28180311