Promoter analysis of the nuclear gene encoding the chloroplast glyceraldehyde-3-phosphate dehydrogenase B subunit of Arabidopsis thaliana

Promoter analysis of the nuclear gene encoding the chloroplast glyceraldehyde-3-phosphate... The promoter of the nuclear gene, GAPB, which encodes the B subunit of chloroplast glyceraldehyde-3-phosphate dehydrogenase (GADPH) of Arabidopsis thaliana, was previously shown to contain four direct repeats (Gap boxes, located between −237 and −181) that were necessary but not sufficient for light-activated gene transcription. To identify additional elements located between the Gap boxes and TATA box, various GAPB promoter fragments driving the β-glucuronidase (GUS) reporter gene were constructed in transgenic Arabidopsis. We found a 23 bp element (the XXIII element), centered at −119, that is essential for promoter activity. Mutations in the XXIII element abolished transcription of GAPB completely. Furthermore, we have identified three additional elements, PI, Tboxes, and PII that serve as positive modulators in the light-activated transcription of GAPB. Mutations in any of these three elements resulted in the reduction in light inducibility of the GAPB gene. The PI, XXIII, Tboxes and PII sequences are novel cis-acting elements that are not present in the closely related GAPA promoter or other promoters that are similarly regulated by light. In our current study, we found that transgenic Arabidopsis containing a GAPB promoter::GUS construct with all four Gap boxes deleted exhibited significant GUS expression albeit reduced to 42% of the optimal expression level. In contrast, in previous studies on transgenic tobacco, total abolishment of GUS expression was seen when the Gap boxes were deleted. This suggests that different trans-acting factors present in heterologous systems may result in variability of the expression of the transgene. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Promoter analysis of the nuclear gene encoding the chloroplast glyceraldehyde-3-phosphate dehydrogenase B subunit of Arabidopsis thaliana

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Publisher
Springer Journals
Copyright
Copyright © 2001 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:1010602031070
Publisher site
See Article on Publisher Site

Abstract

The promoter of the nuclear gene, GAPB, which encodes the B subunit of chloroplast glyceraldehyde-3-phosphate dehydrogenase (GADPH) of Arabidopsis thaliana, was previously shown to contain four direct repeats (Gap boxes, located between −237 and −181) that were necessary but not sufficient for light-activated gene transcription. To identify additional elements located between the Gap boxes and TATA box, various GAPB promoter fragments driving the β-glucuronidase (GUS) reporter gene were constructed in transgenic Arabidopsis. We found a 23 bp element (the XXIII element), centered at −119, that is essential for promoter activity. Mutations in the XXIII element abolished transcription of GAPB completely. Furthermore, we have identified three additional elements, PI, Tboxes, and PII that serve as positive modulators in the light-activated transcription of GAPB. Mutations in any of these three elements resulted in the reduction in light inducibility of the GAPB gene. The PI, XXIII, Tboxes and PII sequences are novel cis-acting elements that are not present in the closely related GAPA promoter or other promoters that are similarly regulated by light. In our current study, we found that transgenic Arabidopsis containing a GAPB promoter::GUS construct with all four Gap boxes deleted exhibited significant GUS expression albeit reduced to 42% of the optimal expression level. In contrast, in previous studies on transgenic tobacco, total abolishment of GUS expression was seen when the Gap boxes were deleted. This suggests that different trans-acting factors present in heterologous systems may result in variability of the expression of the transgene.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 3, 2004

References

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