Minimal sequence requirements for the regulated expression of rbcS-3A from Pisum sativum in transgenic tobacco plants

Cuozzo-Davis, Maria, Yong, Mun-Heng, Gilmartin, Philip M., Goyvaerts, Elisabeth, Kuhlemeier, Cris, Sarokin, Laura and Chua, Nam-Hai (1990) Minimal sequence requirements for the regulated expression of rbcS-3A from Pisum sativum in transgenic tobacco plants. Photochemistry and Photobiology, 52 (1). pp. 43-50. ISSN 0031-8655

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RbcS-3A, the most highly expressed member of the pea multigene family encoding the small subunit of ribulose 1,5-bisphosphate carboxylase, is expressed in a light-dependent and organ-specific manner. In order to further delineate the sequences which mediate this complex pattern of regulation, putative regulatory sequences were assayed for function in transgenic tobacco plants in the context of an inactive 5' deleted rbcS-3A test gene. We have identified a minimal functional unit of 58 bp which is able to confer organ-specific transcriptional activity. It contains two sequences conserved among the pea rbcS family members, namely box II (-151 to -138; GTGTGGTTAATATG) and box III (-125 to -114; ATCATTTTCACT). These sequences bind the nuclear factor termed GT-1 in vitro. Substitution mutations within this 58 bp element have demonstrated that sequences upstream of, or located between, boxes II and III are not required for the transcriptional activity conferred by this element. Distance and orientation of these sequences from the gene are not critical for activity within the limits tested. DNA fragments upstream of nucleotide -170 of rbcS-3A that contain other GT-1 binding sites can also confer regulated expression upon the rbcS-3A promoter deleted to -50. Multimers of individual motifs, namely four tandem copies of boxes II and III, are unable to drive expression of the deleted promoter. These observations suggest that while GT-1 binding is necessary for promoter activity it is by itself not sufficient.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Science > Research Groups > Plant Sciences
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Depositing User: Pure Connector
Date Deposited: 01 Nov 2013 14:28
Last Modified: 24 Oct 2022 04:56

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