Interactions across exons can influence splice site recognition in plant nuclei

In vivo analyses of cis-acting sequence requirements for pre mRNA splicing in tobacco nuclei have previously demonstrated that the 5' splice sites are selected by their position relative to AU-rich elements within plant introns and by their degree of complementarily to the U1 small nuclear RNA....

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Bibliographic Details
Published in:The Plant cell 1996-12, Vol.8 (12), p.2295-2307
Main Authors: McCullough, A.J. (Baylor College of Medicine, Houston, TX.), Baynton, C.E, Schuler, M.A
Format: Article
Language:English
Subjects:
ADN RECOMBINADO
ADN RECOMBINE
Antigens, Plant
ARN MENSAJERO
ARN MESSAGER
Base Sequence
Cell Nucleus - metabolism
Exons
Genetic mutation
Globulins - biosynthesis
Globulins - genetics
GLYCINE MAX
Introns
Molecular Sequence Data
MUTACION
MUTATION
Nicotiana - metabolism
Nicotiana tabacum
Nucleotides
Plant cells
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
Plants, Toxic
Recombinant Fusion Proteins - biosynthesis
Reverse transcriptase polymerase chain reaction
RNA Precursors - metabolism
RNA Splicing
RNA, Plant - metabolism
RNA, Small Nuclear - chemistry
RUBISCO
Seed Storage Proteins
Site selection
Small nuclear RNA
Soybean Proteins
Splicing
Transcription, Genetic
Transfection
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Summary:In vivo analyses of cis-acting sequence requirements for pre mRNA splicing in tobacco nuclei have previously demonstrated that the 5' splice sites are selected by their position relative to AU-rich elements within plant introns and by their degree of complementarily to the U1 small nuclear RNA. To determine whether the presence of adjacent introns affects 5' splice site recognition in plant nuclei, we have analyzed the in vivo spiking patterns of two-intron constructs containing 5' splice site mutations in the second intron. These experiments indicated that the splice site selection patterns in plant nuclei are defined primarily by sequences within the Intron (Intron definition) and secondarily by weak interactions across exons (exon definition). The effects of these secondary interactions became evident only when mutations in the downstream 5' splice site decreased its functionality and differed depending on the availability of cryptic splice sites close to the mutant site. In beta-conglycinin chimeric transcripts containing multiple cryptic 5' splice sites, the presence of an intact upstream intron significantly increased splicing at the downstream 5' splice sites in a polar fashion without activating exon skipping. In a natural 3-conglycinin transcript, which does not contain cryptic 5' splice sites, mutation of the first nucleotide of the downstream intron activated an array of noncanonical 5' end 3' splice sites and some exon skipping
ISSN:1040-4651
1532-298X
DOI:10.1105/tpc.8.12.2295