Heterotrimeric G proteins are implicated in gibberellin induction of α-amylase gene expression in wild oat aleurone

H D Jones, S J Smith, R Desikan, S Plakidou-Dymock, A Lovegrove, Richard Hooley

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Abstract

The role of heterotrimeric G proteins in gibberellin (GA) induction of α-amylase gene expression was examined in wild oat aleurone protoplasts. Mas7, a cationic amphiphilic tetradecapeptide that stimulates GDP/GTP exchange by heterotrimeric G proteins, specifically induced alpha-amylase gene expression and enzyme secretion in a very similar manner to GA(1). In addition, Mas7 stimulated expression of an alpha-Amy2/54:GUS promoter and reporter construct in transformed protoplasts. Both Mas7 and GA(1) induction of α-amylase mRNA were insensitive to pertussis toxin. Hydrolysis-resistant nucleotides were introduced into aleurone protoplasts during transfection with reporter gene constructs. GDP-beta-S, which inhibits GDP/GTP exchange by heterotrimeric G proteins, completely prevented GA(1) induction of α-Amy2/ 54:GUS expression, whereas GTP-gamma-S, which activates heterotrimeric G proteins, stimulated expression very slightly. Novel cDNA sequences from G(alpha) and G(beta) subunits were cloned from wild oat aleurone cells. By using RNA gel blot analysis, we found that the transcripts were expressed at a low level. Heterotrimeric G proteins have been implicated in several events during plant growth and development, and these data suggest that they may be involved in GA regulation of α-amylase gene expression in aleurone.
LanguageEnglish
Pages245-253
Number of pages9
JournalPlant Cell
Volume10
Issue number2
StatusPublished - 1998

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Gibberellins
Heterotrimeric GTP-Binding Proteins
alpha-Amylases
G-proteins
alpha-amylase
gibberellins
oats
Protoplasts
Gene Expression
gene expression
protoplasts
Guanosine Triphosphate
aleurone cells
Guanosine 5'-O-(3-Thiotriphosphate)
pertussis toxin
Plant Development
Pertussis Toxin
transfection
Reporter Genes
Growth and Development

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Heterotrimeric G proteins are implicated in gibberellin induction of α-amylase gene expression in wild oat aleurone. / Jones, H D; Smith, S J; Desikan, R; Plakidou-Dymock, S; Lovegrove, A; Hooley, Richard.

In: Plant Cell, Vol. 10, No. 2, 1998, p. 245-253.

Research output: Contribution to journalArticle

Jones, HD, Smith, SJ, Desikan, R, Plakidou-Dymock, S, Lovegrove, A & Hooley, R 1998, 'Heterotrimeric G proteins are implicated in gibberellin induction of α-amylase gene expression in wild oat aleurone' Plant Cell, vol. 10, no. 2, pp. 245-253.
Jones, H D ; Smith, S J ; Desikan, R ; Plakidou-Dymock, S ; Lovegrove, A ; Hooley, Richard. / Heterotrimeric G proteins are implicated in gibberellin induction of α-amylase gene expression in wild oat aleurone. In: Plant Cell. 1998 ; Vol. 10, No. 2. pp. 245-253
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abstract = "The role of heterotrimeric G proteins in gibberellin (GA) induction of α-amylase gene expression was examined in wild oat aleurone protoplasts. Mas7, a cationic amphiphilic tetradecapeptide that stimulates GDP/GTP exchange by heterotrimeric G proteins, specifically induced alpha-amylase gene expression and enzyme secretion in a very similar manner to GA(1). In addition, Mas7 stimulated expression of an alpha-Amy2/54:GUS promoter and reporter construct in transformed protoplasts. Both Mas7 and GA(1) induction of α-amylase mRNA were insensitive to pertussis toxin. Hydrolysis-resistant nucleotides were introduced into aleurone protoplasts during transfection with reporter gene constructs. GDP-beta-S, which inhibits GDP/GTP exchange by heterotrimeric G proteins, completely prevented GA(1) induction of α-Amy2/ 54:GUS expression, whereas GTP-gamma-S, which activates heterotrimeric G proteins, stimulated expression very slightly. Novel cDNA sequences from G(alpha) and G(beta) subunits were cloned from wild oat aleurone cells. By using RNA gel blot analysis, we found that the transcripts were expressed at a low level. Heterotrimeric G proteins have been implicated in several events during plant growth and development, and these data suggest that they may be involved in GA regulation of α-amylase gene expression in aleurone.",
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N2 - The role of heterotrimeric G proteins in gibberellin (GA) induction of α-amylase gene expression was examined in wild oat aleurone protoplasts. Mas7, a cationic amphiphilic tetradecapeptide that stimulates GDP/GTP exchange by heterotrimeric G proteins, specifically induced alpha-amylase gene expression and enzyme secretion in a very similar manner to GA(1). In addition, Mas7 stimulated expression of an alpha-Amy2/54:GUS promoter and reporter construct in transformed protoplasts. Both Mas7 and GA(1) induction of α-amylase mRNA were insensitive to pertussis toxin. Hydrolysis-resistant nucleotides were introduced into aleurone protoplasts during transfection with reporter gene constructs. GDP-beta-S, which inhibits GDP/GTP exchange by heterotrimeric G proteins, completely prevented GA(1) induction of α-Amy2/ 54:GUS expression, whereas GTP-gamma-S, which activates heterotrimeric G proteins, stimulated expression very slightly. Novel cDNA sequences from G(alpha) and G(beta) subunits were cloned from wild oat aleurone cells. By using RNA gel blot analysis, we found that the transcripts were expressed at a low level. Heterotrimeric G proteins have been implicated in several events during plant growth and development, and these data suggest that they may be involved in GA regulation of α-amylase gene expression in aleurone.

AB - The role of heterotrimeric G proteins in gibberellin (GA) induction of α-amylase gene expression was examined in wild oat aleurone protoplasts. Mas7, a cationic amphiphilic tetradecapeptide that stimulates GDP/GTP exchange by heterotrimeric G proteins, specifically induced alpha-amylase gene expression and enzyme secretion in a very similar manner to GA(1). In addition, Mas7 stimulated expression of an alpha-Amy2/54:GUS promoter and reporter construct in transformed protoplasts. Both Mas7 and GA(1) induction of α-amylase mRNA were insensitive to pertussis toxin. Hydrolysis-resistant nucleotides were introduced into aleurone protoplasts during transfection with reporter gene constructs. GDP-beta-S, which inhibits GDP/GTP exchange by heterotrimeric G proteins, completely prevented GA(1) induction of α-Amy2/ 54:GUS expression, whereas GTP-gamma-S, which activates heterotrimeric G proteins, stimulated expression very slightly. Novel cDNA sequences from G(alpha) and G(beta) subunits were cloned from wild oat aleurone cells. By using RNA gel blot analysis, we found that the transcripts were expressed at a low level. Heterotrimeric G proteins have been implicated in several events during plant growth and development, and these data suggest that they may be involved in GA regulation of α-amylase gene expression in aleurone.

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