Identification of cis-regulatory elements required for endosperm expression of the rice storage protein glutelin gene GluB-1

Identification of cis-regulatory elements required for endosperm expression of the rice storage... Rice storage protein glutelin genes are coordinately regulated during seed development. A previous 5′ deletion analysis using transgenic tobacco revealed that the minimum 5′ region necessary for endosperm specificity was within −245 bp of the transcription start site, and included the AACA and GCN4 motifs that are highly conserved in the 5′-flanking regions of all glutelin genes. In this paper, the sequence elements essential for endosperm-specific expression are characterized in stable transgenic tobacco plants by both loss-of-function and gain-of-function experiments using this minimum promoter. Base substitution analysis shows that the proximal AACA motif between −73 and −61, and the GCN4 motif between −165 and −158 act as critical elements. An ACGT motif between −81 and −75, and Skn-I-like elements between −173 and −169 also play important roles in controlling the seed-specific expression. When the distal region between −245 and −145 containing the AACA and the GCN4 motifs or the proximal region between −113 and −46 containing the ACGT and AACA motifs is fused to a truncated promoter (−90 to +9) of the CaMV 35S gene fused to the β-glucuronidase (GUS) reporter gene, high levels of seed-specific expression are observed in these fusions, thereby indicating that either pair of motifs is sufficient to confer seed expression in these fusions. However, when substituted for by the CaMV 35S core promoter (−46 to +1), seed expression is abolished, suggesting that the sequence between −90 and −46 of the CaMV 35S promoter containing G-box-like motif (as-1 element) is required for such specific expression in addition to AACA and GCN4 motifs. Therefore, we conclude that at least three cis-regulatory elements, the AACA motif, GCN4 motif and ACGT motif, are necessary to mediate endosperm expression of the GluB-1 glutelin gene. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Identification of cis-regulatory elements required for endosperm expression of the rice storage protein glutelin gene GluB-1

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 1999 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1026459229671
Publisher site
See Article on Publisher Site

Abstract

Rice storage protein glutelin genes are coordinately regulated during seed development. A previous 5′ deletion analysis using transgenic tobacco revealed that the minimum 5′ region necessary for endosperm specificity was within −245 bp of the transcription start site, and included the AACA and GCN4 motifs that are highly conserved in the 5′-flanking regions of all glutelin genes. In this paper, the sequence elements essential for endosperm-specific expression are characterized in stable transgenic tobacco plants by both loss-of-function and gain-of-function experiments using this minimum promoter. Base substitution analysis shows that the proximal AACA motif between −73 and −61, and the GCN4 motif between −165 and −158 act as critical elements. An ACGT motif between −81 and −75, and Skn-I-like elements between −173 and −169 also play important roles in controlling the seed-specific expression. When the distal region between −245 and −145 containing the AACA and the GCN4 motifs or the proximal region between −113 and −46 containing the ACGT and AACA motifs is fused to a truncated promoter (−90 to +9) of the CaMV 35S gene fused to the β-glucuronidase (GUS) reporter gene, high levels of seed-specific expression are observed in these fusions, thereby indicating that either pair of motifs is sufficient to confer seed expression in these fusions. However, when substituted for by the CaMV 35S core promoter (−46 to +1), seed expression is abolished, suggesting that the sequence between −90 and −46 of the CaMV 35S promoter containing G-box-like motif (as-1 element) is required for such specific expression in addition to AACA and GCN4 motifs. Therefore, we conclude that at least three cis-regulatory elements, the AACA motif, GCN4 motif and ACGT motif, are necessary to mediate endosperm expression of the GluB-1 glutelin gene.

Journal

Plant Molecular BiologySpringer Journals

Published: Sep 29, 2004

References

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