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Delayed Trans-inactivation of Synthetic Domain A 35S Promoters by “Tobacco 271 Locus” due to Reduced Sequence Homology

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Abstract

Homology in the promoter region leads to transcriptional gene silencing (TGS). This could be circumvented by designing synthetic promoters which are functionally similar but divergent in their sequence. In the present study, we asked whether such synthetic promoters can escape trans-inactivation when challenged with a silenced copy of the wild-type prototype. We tested the activity of two synthetic cauliflower mosaic virus 35S (35S) domain A promoters, 2mA and 3mA, with only 35% and 41% sequence identity with the wild type 35S (wtA) in the presence of silencing locus 271 of Nicotiana tabacum that inactivates any 35S promoter driven transgene through TGS. Based on comparison of β-glucuronidase (gus) activity under the transcriptional control of wtA, 2mA, and 3mA in the presence and absence of the silencing locus, we demonstrate that the inactivation of the 2mA and 3mA promoters is significantly delayed in comparison to that of wtA although microhomology at the transcription factor binding sites eventually leads to the silencing of these synthetic promoters. We further postulate that it is possible to design functional synthetic promoters that can escape gene silencing in the background of silencing locus if the cis-elements are smaller and do not contain methylation prone CG and CNG.

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Acknowledgment

We thank Dr. Herve Vaucheret, INRA Centre de Versailles, France, for providing us the seed material for silencing line 271.5.8 and nonsilencing line 271.5.9. We thank Ms. Taru Gautam for her technical assistance and Dr. Anjana N. Dev for critically going through the manuscript. This work was supported by grants from Department of Biotechnology, Government of India and University of Delhi, India. SB and SD were supported by Research Fellowships from Council of Scientific and Industrial Research, Government of India.

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  1. Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Simran Bhullar, Sudipta Datta & Pradeep Kumar Burma

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  1. Simran Bhullar
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  2. Sudipta Datta
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  3. Pradeep Kumar Burma
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Correspondence to Pradeep Kumar Burma.

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Simran Bhullar and Sudipta Datta contributed equally to this study.

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Bhullar, S., Datta, S. & Burma, P.K. Delayed Trans-inactivation of Synthetic Domain A 35S Promoters by “Tobacco 271 Locus” due to Reduced Sequence Homology. Plant Mol Biol Rep 29, 1–11 (2011). https://doi.org/10.1007/s11105-010-0202-4

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