hnRNP H enhances skipping of a nonfunctional exon P3A in CHRNA1 and a mutation disrupting its binding causes congenital myasthenic syndrome

Akio Masuda, Xin Ming Shen, Mikako Ito, Tohru Matsuura, Andrew G Engel, Kinji Ohno

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

In humans and great apes, CHRNA1 encoding the muscle nicotinic acetylcholine receptor α subunit carries an inframe exon P3A, the inclusion of which yields a nonfunctional α subunit. In muscle, the P3A(-) and P3A(+) transcripts are generated in a 1:1 ratio but the functional significance and regulation of the alternative splicing remain elusive. An intronic mutation (IVS3-8G>A), identified in a patient with congenital myasthenic syndrome, disrupts an intronic splicing silencer (ISS) and results in exclusive inclusion of the downstream P3A exon. We found that the ISS-binding splicing trans -factor was heterogeneous nuclear ribonucleoprotein (hnRNP) H and the mutation attenuated the affinity of hnRNP for the ISS ∼100-fold. We next showed that direct placement of hnRNP H to the 3′ end of intron 3 silences, and siRNA-mediated downregulation of hnRNP H enhances recognition of exon P3A. Analysis of the human genome suggested that the hnRNPH-binding UGGG motif is overrepresented close to the 3′ ends of introns. Pursuing this clue, we showed that alternative exons of GRIP1, FAS, VPS13C and NRCAM are downregulated by hnRNP H. Our findings imply that the presence of the hnRNP H-binding motif close to the 3′ end of an intron is an essential but underestimated splicing regulator of the downstream exon.

Original languageEnglish (US)
Pages (from-to)4022-4035
Number of pages14
JournalHuman Molecular Genetics
Volume17
Issue number24
DOIs
StatePublished - 2008

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Congenital Myasthenic Syndromes
Heterogeneous-Nuclear Ribonucleoproteins
Exons
Mutation
Introns
Down-Regulation
Muscles
Hominidae
Alternative Splicing
Nicotinic Receptors
Human Genome
Small Interfering RNA

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Genetics(clinical)

Cite this

hnRNP H enhances skipping of a nonfunctional exon P3A in CHRNA1 and a mutation disrupting its binding causes congenital myasthenic syndrome. / Masuda, Akio; Shen, Xin Ming; Ito, Mikako; Matsuura, Tohru; Engel, Andrew G; Ohno, Kinji.

In: Human Molecular Genetics, Vol. 17, No. 24, 2008, p. 4022-4035.

Research output: Contribution to journalArticle

Masuda, Akio ; Shen, Xin Ming ; Ito, Mikako ; Matsuura, Tohru ; Engel, Andrew G ; Ohno, Kinji. / hnRNP H enhances skipping of a nonfunctional exon P3A in CHRNA1 and a mutation disrupting its binding causes congenital myasthenic syndrome. In: Human Molecular Genetics. 2008 ; Vol. 17, No. 24. pp. 4022-4035.
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