title: Transcriptional expression of cis-acting and trans-acting splicing mutations cause autosomal dominant retinitis pigmentosa.
abstract: Two types of mutations may lead to deficient pre-mRNA splicing: cis-acting mutations that inactivate a constitutive or alternative splice site within the pre-mRNA, and trans-acting mutations that affect the function of a basal factor of the splicing machinery. Autosomal dominant retinitis pigmentosa (adRP) is caused by mutations in at least 12 genes, with mutations in rhodopsin being the most prevalent. Two cis-acting mutations, g.3811A>G and g.5167G>T at the splice site in the rhodopsin gene (RHO; GenBank U49742.1) are linked to adRP in a Spanish population; while a cis-acting mutation, g.4335G>T, has been linked to recessive RP (arRP). Transcriptional expression analysis showed that the cis-acting splicing mutations linked to adRP promoted alternative splice sites, while the arRP linked mutation results in exclusion of exon 4. Trans-acting splicing mutations associated with adRP have also been found, and mutations in the pre-mRNA splicing factors PRPF3, PRPF8, PRPF31, and RP9 are associated with adRP in several populations. This report describes a new mutation in PRPF3 in a Spanish adRP family. We also investigated the transcriptional patterns in Epstein-Barr virus (EBV)-transformed lymphoblastoid cells from patients carrying a mutation in PRPF8. Despite the role of PRPF8 in the minor U12 splicing processes, microarray analysis revealed that mutations in PRPF8 not only did not result in significant differences in splicing efficiency of rhodopsin, but no apparent changes in expression of U12-type intron genes and splicing processes was observed. Microarray analysis revealed a panel of differentially expressed genes mapped to the RP loci, and future work will determine their role in RP.
authors.affiliation: Servicio de Laboratorio, Departamento de Genética y Biología Molecular, Hospital de Terrassa, Terrassa, Barcelona, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: Gamundi
authors.forename: Imma
authors.initials: I
authors.lastname: Hernan
authors.forename: Marta
authors.initials: M
authors.lastname: Muntanyola
authors.forename: Miquel
authors.initials: M
authors.lastname: Maseras
authors.forename: Pedro
authors.initials: P
authors.lastname: López-Romero
authors.forename: Rebeca
authors.initials: R
authors.lastname: Alvarez
authors.forename: Ana
authors.initials: A
authors.lastname: Dopazo
authors.forename: Salud
authors.initials: S
authors.lastname: Borrego
authors.forename: Miguel
authors.initials: M
authors.lastname: Carballo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D004252:DNA Mutational Analysis; D005260:Female; D005799:Genes, Dominant; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010375:Pedigree; D012326:RNA Splicing; D016601:RNA-Binding Proteins; D012174:Retinitis Pigmentosa / Q000235:genetics*; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013030:Spain
chemical_list: D002352:Carrier Proteins; D009687:Nuclear Proteins; C108507:PRPF3 protein, human; C115374:PRPF8 protein, human; D016601:RNA-Binding Proteins; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; D012243:Rhodopsin