title: Structural determinants for alternative splicing regulation of the MAPT pre-mRNA.
abstract: Alternative splicing at the MAPT gene exon 10 yields similar levels of the 3R and 4R tau protein isoforms. (1) The presence of mutations, particularly in exon 10 and intron 10-11, changes the quantity of tau isoforms. Domination each of the isoform yields tau protein aggregation and frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17). Here, we report for the first time the secondary structure of the 194/195 nucleotide region for the wild type (WT) and 10 mutants of the MAPT gene pre-mRNA determined using both chemical and microarray mapping. Thermodynamic analyses indicate that single nucleotide mutations in the splicing regulatory element (SRE) that form a hairpin affect its stability by up to 4 and 7 kcal/mol. Moreover, binding the regulatory hairpin of small molecule ligands (neomycin, kanamycin, tobramycin and mitoxantrone) enhance its stability depending on the nature of the ligands and the RNA mutations. Experiments using the cos-7 cell line indicate that the presence of ligands and modified antisense oligonucleotides affect the quantity of 3R and 4R isoforms. This finding correlates with the thermodynamic stability of the regulatory hairpin. An alternative splicing regulation mechanism for exon 10 is postulated based on our experimental data and on published data.
authors.affiliation: a Institute of Bioorganic Chemistry; Polish Academy of Sciences ; Noskowskiego, Poland.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Lisowiec
authors.forename: Dorota
authors.initials: D
authors.lastname: Magner
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Kierzek
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Lenartowicz
authors.forename: Ryszard
authors.initials: R
authors.lastname: Kierzek
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D004742:Enhancer Elements, Genetic; D005091:Exons; D006801:Humans; D007438:Introns; D008024:Ligands; D046228:Microarray Analysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry; D012322:RNA Precursors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D037941:Silencer Elements, Transcriptional; D013816:Thermodynamics; D016875:tau Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D008024:Ligands; C054369:MAPT protein, human; D016376:Oligonucleotides, Antisense; D012322:RNA Precursors; D011994:Recombinant Proteins; D016875:tau Proteins
keywords: AD, Alzheimer disease; DMS, dimethyl sulfide; ESE, exonic splicing enhancer; ESS, exonic splicing silencer; FTD, frontotemporal dementia; FTDP-17, frontotemporal dementia and Parkinsonism linked to chromosome 17; ISM, intronic splicing modulator; ISS, intronic splicing silencer; MAPT, microtubule-associated protein tau; NMIA, N-methylisotoic anhydride; NMR, nuclear magnetic resonance; PPE, polypurine enhancer; RNA structure; RNA thermodynamics; RT-PCR, reverse transcription polymerase chain reaction; SHAPE, selective 2′-hydroxyl acylation analyzed by primer extension; SMA, spinal muscular atrophy; SRE, splicing regulatory element; U1 snRNP, U1 small nuclear ribonucleoprotein; WT, wild type; alternative splicing regulation; antisense oligonucleotides; neurodegradation; pre-mRNA, pre-messenger RNA; small molecule binding
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