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Differential 3' splice site recognition of SMN1 and SMN2 transcripts by U2AF and U2 snRNP.
RNA. 2009 Apr; 15(4):515-23.RNA

Abstract

Spinal Muscular atrophy is a prevalent genetic disease caused by mutation of the SMN1 gene, which encodes the SMN protein involved in assembly of small nuclear ribonucleoprotein (snRNP) complexes. A paralog of the gene, SMN2, cannot provide adequate levels of functional SMN because exon 7 is skipped in a significant fraction of the mature transcripts. A C to T transition located at position 6 of exon 7 is critical for the difference in exon skipping between SMN1 and SMN2. Here we report that this nucleotide difference results in increased ultraviolet light-mediated crosslinking of the splicing factor U2AF(65) with the 3' splice site of SMN1 intron 6 in HeLa nuclear extract. U2 snRNP association, analyzed by native gel electrophoresis, is also more efficient on SMN1 than on SMN2, particularly under conditions of competition, suggesting more effective use of limiting factors. Two trans-acting factors implicated in SMN regulation, SF2/ASF and hnRNP A1, promote and repress, respectively, U2 snRNP recruitment to both RNAs. Interestingly, depending on the transcript and the regulatory factor, the effects on U2 binding not always correlate with changes in U2AF(65) crosslinking. Furthermore, blocking recognition of a Tra2-beta1-dependent splicing enhancer located in exon 7 inhibits U2 snRNP recruitment without affecting U2AF(65) crosslinking. Collectively, the results suggest that both U2AF binding and other steps of U2 snRNP recruitment can be control points in SMN splicing regulation.

Authors+Show Affiliations

Centre de Regulació Genòmica, Barcelona, Spain.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19244360

Citation

Martins de Araújo, Mafalda, et al. "Differential 3' Splice Site Recognition of SMN1 and SMN2 Transcripts By U2AF and U2 SnRNP." RNA (New York, N.Y.), vol. 15, no. 4, 2009, pp. 515-23.
Martins de Araújo M, Bonnal S, Hastings ML, et al. Differential 3' splice site recognition of SMN1 and SMN2 transcripts by U2AF and U2 snRNP. RNA. 2009;15(4):515-23.
Martins de Araújo, M., Bonnal, S., Hastings, M. L., Krainer, A. R., & Valcárcel, J. (2009). Differential 3' splice site recognition of SMN1 and SMN2 transcripts by U2AF and U2 snRNP. RNA (New York, N.Y.), 15(4), 515-23. https://doi.org/10.1261/rna.1273209
Martins de Araújo M, et al. Differential 3' Splice Site Recognition of SMN1 and SMN2 Transcripts By U2AF and U2 SnRNP. RNA. 2009;15(4):515-23. PubMed PMID: 19244360.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Differential 3' splice site recognition of SMN1 and SMN2 transcripts by U2AF and U2 snRNP. AU - Martins de Araújo,Mafalda, AU - Bonnal,Sophie, AU - Hastings,Michelle L, AU - Krainer,Adrian R, AU - Valcárcel,Juan, Y1 - 2009/02/25/ PY - 2009/2/27/entrez PY - 2009/2/27/pubmed PY - 2009/4/2/medline SP - 515 EP - 23 JF - RNA (New York, N.Y.) JO - RNA VL - 15 IS - 4 N2 - Spinal Muscular atrophy is a prevalent genetic disease caused by mutation of the SMN1 gene, which encodes the SMN protein involved in assembly of small nuclear ribonucleoprotein (snRNP) complexes. A paralog of the gene, SMN2, cannot provide adequate levels of functional SMN because exon 7 is skipped in a significant fraction of the mature transcripts. A C to T transition located at position 6 of exon 7 is critical for the difference in exon skipping between SMN1 and SMN2. Here we report that this nucleotide difference results in increased ultraviolet light-mediated crosslinking of the splicing factor U2AF(65) with the 3' splice site of SMN1 intron 6 in HeLa nuclear extract. U2 snRNP association, analyzed by native gel electrophoresis, is also more efficient on SMN1 than on SMN2, particularly under conditions of competition, suggesting more effective use of limiting factors. Two trans-acting factors implicated in SMN regulation, SF2/ASF and hnRNP A1, promote and repress, respectively, U2 snRNP recruitment to both RNAs. Interestingly, depending on the transcript and the regulatory factor, the effects on U2 binding not always correlate with changes in U2AF(65) crosslinking. Furthermore, blocking recognition of a Tra2-beta1-dependent splicing enhancer located in exon 7 inhibits U2 snRNP recruitment without affecting U2AF(65) crosslinking. Collectively, the results suggest that both U2AF binding and other steps of U2 snRNP recruitment can be control points in SMN splicing regulation. SN - 1469-9001 UR - https://www.unboundmedicine.com/medline/citation/19244360/Differential_3'_splice_site_recognition_of_SMN1_and_SMN2_transcripts_by_U2AF_and_U2_snRNP_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=19244360 DB - PRIME DP - Unbound Medicine ER -