Direct evidence for rapid degradation of bacillus thuringiensis toxin mRNA as a cause of poor expression in plants

E. Jay De Rocher, Tracy Vargo-Gogola, Scott H. Diehn, Pamela J. Green

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

It is well established that the expression of Bacillus thuringiensis (B.t.) toxin genes in higher plants is severely limited at the mRNA level, but the cause remains controversial. Elucidating whether mRNA accumulation is limited transcriptionally or posttranscriptionally could contribute to effective gene design as well as provide insights about endogenous plant gene-expression mechanisms. To resolve this controversy, we compared the expression of an A/U-rich wild-type cryIA(c) gene and a G/C-rich synthetic cryIA(c) B.t.-toxin gene under the control of identical 5′ and 3′ flanking sequences. Transcriplional activities of the genes were equal as determined by nuclear run-on transcription assays. In contrast, mRNA half-life measurements demonstrated directly that the wildtype transcript was markedly less stable than that encoded by the synthetic gene. Sequences that limit mRNA accumulation were located at more than one site within the coding region, and some appeared to be recognized in Arabidopsis but not in tobacco (Nicotiana tabacum). These results support previous observations that some A/U-rich sequences can contribute to mRNA instability in plants. Our studies further indicate that some of these sequences may be differentially recognized in tobacco cells and Arabidopsis.

Original languageEnglish (US)
Pages (from-to)1445-1461
Number of pages17
JournalPlant Physiology
Volume117
Issue number4
StatePublished - 1998
Externally publishedYes

Fingerprint

Bacillus thuringiensis
toxins
Messenger RNA
degradation
Tobacco
Genes
genes
Arabidopsis
tobacco
3' Flanking Region
Synthetic Genes
synthetic genes
Plant Genes
5' Flanking Region
RNA Stability
Nicotiana tabacum
half life
Half-Life
transcription (genetics)
Gene Expression

ASJC Scopus subject areas

  • Plant Science

Cite this

Direct evidence for rapid degradation of bacillus thuringiensis toxin mRNA as a cause of poor expression in plants. / Jay De Rocher, E.; Vargo-Gogola, Tracy; Diehn, Scott H.; Green, Pamela J.

In: Plant Physiology, Vol. 117, No. 4, 1998, p. 1445-1461.

Research output: Contribution to journalArticle

Jay De Rocher, E. ; Vargo-Gogola, Tracy ; Diehn, Scott H. ; Green, Pamela J. / Direct evidence for rapid degradation of bacillus thuringiensis toxin mRNA as a cause of poor expression in plants. In: Plant Physiology. 1998 ; Vol. 117, No. 4. pp. 1445-1461.
@article{41ccabbcaeac4a09bc7a66371fea9695,
title = "Direct evidence for rapid degradation of bacillus thuringiensis toxin mRNA as a cause of poor expression in plants",
abstract = "It is well established that the expression of Bacillus thuringiensis (B.t.) toxin genes in higher plants is severely limited at the mRNA level, but the cause remains controversial. Elucidating whether mRNA accumulation is limited transcriptionally or posttranscriptionally could contribute to effective gene design as well as provide insights about endogenous plant gene-expression mechanisms. To resolve this controversy, we compared the expression of an A/U-rich wild-type cryIA(c) gene and a G/C-rich synthetic cryIA(c) B.t.-toxin gene under the control of identical 5′ and 3′ flanking sequences. Transcriplional activities of the genes were equal as determined by nuclear run-on transcription assays. In contrast, mRNA half-life measurements demonstrated directly that the wildtype transcript was markedly less stable than that encoded by the synthetic gene. Sequences that limit mRNA accumulation were located at more than one site within the coding region, and some appeared to be recognized in Arabidopsis but not in tobacco (Nicotiana tabacum). These results support previous observations that some A/U-rich sequences can contribute to mRNA instability in plants. Our studies further indicate that some of these sequences may be differentially recognized in tobacco cells and Arabidopsis.",
author = "{Jay De Rocher}, E. and Tracy Vargo-Gogola and Diehn, {Scott H.} and Green, {Pamela J.}",
year = "1998",
language = "English (US)",
volume = "117",
pages = "1445--1461",
journal = "Plant Physiology",
issn = "0032-0889",
publisher = "American Society of Plant Biologists",
number = "4",

}

TY - JOUR

T1 - Direct evidence for rapid degradation of bacillus thuringiensis toxin mRNA as a cause of poor expression in plants

AU - Jay De Rocher, E.

AU - Vargo-Gogola, Tracy

AU - Diehn, Scott H.

AU - Green, Pamela J.

PY - 1998

Y1 - 1998

N2 - It is well established that the expression of Bacillus thuringiensis (B.t.) toxin genes in higher plants is severely limited at the mRNA level, but the cause remains controversial. Elucidating whether mRNA accumulation is limited transcriptionally or posttranscriptionally could contribute to effective gene design as well as provide insights about endogenous plant gene-expression mechanisms. To resolve this controversy, we compared the expression of an A/U-rich wild-type cryIA(c) gene and a G/C-rich synthetic cryIA(c) B.t.-toxin gene under the control of identical 5′ and 3′ flanking sequences. Transcriplional activities of the genes were equal as determined by nuclear run-on transcription assays. In contrast, mRNA half-life measurements demonstrated directly that the wildtype transcript was markedly less stable than that encoded by the synthetic gene. Sequences that limit mRNA accumulation were located at more than one site within the coding region, and some appeared to be recognized in Arabidopsis but not in tobacco (Nicotiana tabacum). These results support previous observations that some A/U-rich sequences can contribute to mRNA instability in plants. Our studies further indicate that some of these sequences may be differentially recognized in tobacco cells and Arabidopsis.

AB - It is well established that the expression of Bacillus thuringiensis (B.t.) toxin genes in higher plants is severely limited at the mRNA level, but the cause remains controversial. Elucidating whether mRNA accumulation is limited transcriptionally or posttranscriptionally could contribute to effective gene design as well as provide insights about endogenous plant gene-expression mechanisms. To resolve this controversy, we compared the expression of an A/U-rich wild-type cryIA(c) gene and a G/C-rich synthetic cryIA(c) B.t.-toxin gene under the control of identical 5′ and 3′ flanking sequences. Transcriplional activities of the genes were equal as determined by nuclear run-on transcription assays. In contrast, mRNA half-life measurements demonstrated directly that the wildtype transcript was markedly less stable than that encoded by the synthetic gene. Sequences that limit mRNA accumulation were located at more than one site within the coding region, and some appeared to be recognized in Arabidopsis but not in tobacco (Nicotiana tabacum). These results support previous observations that some A/U-rich sequences can contribute to mRNA instability in plants. Our studies further indicate that some of these sequences may be differentially recognized in tobacco cells and Arabidopsis.

UR - http://www.scopus.com/inward/record.url?scp=0032134544&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032134544&partnerID=8YFLogxK

M3 - Article

C2 - 9701600

AN - SCOPUS:0032134544

VL - 117

SP - 1445

EP - 1461

JO - Plant Physiology

JF - Plant Physiology

SN - 0032-0889

IS - 4

ER -