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PMID: 12458797 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Requirements for intron-mediated enhancement of gene expression in Arabidopsis.

RNA (New York, N.Y.) ·Vol. 8 ·No. 11 ·2002-11-00 ·Pages 1444-53

Rose AB

Abstract

To explore possible mechanisms of intron-mediated enhancement of gene expression, the features of PAT1 intron 1 required to elevate mRNA accumulation were systematically tested in transgenic Arabidopsis. This intron is remarkably resilient, retaining some ability to increase mRNA accumulation when splicing was prevented by mutation of 5' and 3' splice sites, branchpoint sequences, or when intron U-richness was reduced. Enhancement was abolished by simultaneously eliminating branchpoints and the 5' splice site, structures involved in the first two steps of spliceosome assembly. Although this suggests that the splicing machinery is required, intron splicing is clearly not enough to enhance mRNA accumulation. Five other introns were all efficiently spliced but varied widely in their ability to increase mRNA levels. Furthermore, PAT1 intron 1 was spliced but lost the ability to elevate mRNA accumulation when moved to the 3' UTR. These findings demonstrate that splicing per se is neither necessary nor sufficient for an intron to enhance mRNA accumulation, and suggest a mechanism that requires intron recognition by the splicing machinery but also involves nonconserved intron sequences.

MeSH Terms
Arabidopsis/genetics Base Sequence DNA Primers DNA, Plant Gene Expression Genes, Plant Introns Molecular Sequence Data Mutagenesis Mutation Plants, Genetically Modified/genetics RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA Splice Sites/genetics RNA Splicing RNA, Messenger/metabolism Spliceosomes/metabolism
Chemicals
DNA Primers DNA, Plant RNA Precursors RNA Splice Sites RNA, Messenger
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Rose Alan B
Molecular and Cellular Biology, University of California, Davis, California, 95616, USA. abrose@ucdavis.edu
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s1355838202020551
Email
abrose@ucdavis.edu
Published
2002-11-00
Pages
1444-53
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1370350
Subset
IM
External Links
PubMed source DOI: 10.1017/s1355838202020551 PMC Full Text View journal details
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