Plant Physiology Preview
Published on September 14, 2007; 10.1104/pp.107.104828
Received June 28, 2007
Accepted September 10, 2007
The Transcription Factor RAP2.2 and its Interacting Partner SINAT2: Stable Elements in the Carotenogenesis of Arabidopsis Leaves
Ralf Welsch *, Dirk Maaß , Tanja Voegel , Dean DellaPenna , and Peter Beyer
Faculty of Biology, Center for Applied Biosciences, Universität Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany; Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824, USA
* Corresponding author; email: welschra{at}web.de.
The promoter of phytoene synthase (PSY), the first specific enzyme of carotenoid biosynthesis, shows two main regulatory regions: a G-box containing region located near the TATA box, and a TATA box distal region containing the cis-acting element ATCTA which mediates strong basal promoter activity. This second element was also present in the promoter of phytoene desaturase (PDS), the next step of the carotenoid pathway, suggesting a common regulatory mechanism. In the present work, we demonstrate that AtRAP2.2, a member of the AP2/EREBP transcription factor family, binds to the ATCTA element. In Arabidopsis leaves, AtRAP2.2 transcript and protein levels were tightly controlled as indicated by unchanged transcript and protein levels in T-DNA insertion mutants in the AtRAP2.2 promoter and 5' UTR and the lack of change in AtRAP2.2 protein levels in lines strongly overexpressing the AtRAP2.2 transcript. Homozygous loss-of-function mutants could not be obtained for the AtRAP2.2 5' UTR T-DNA insertion line indicating a lethal phenotype. In AtRAP2.2 overexpression lines, modest changes in PSY and PDS transcripts were only observed in root-derived calli, which consequently showed a reduction in carotenoid content. The RING finger protein "SEVEN IN ABSENTIA OF ARABIDOPSIS 2" (SINAT2) was identified as an AtRAP2.2 interaction partner using a two-hybrid approach. The structure of SINAT2 and related proteins of Arabidopsis show homology to the "SEVEN IN ABSENTIA" protein of Drosophila that is involved in the proteasome-mediated regulation in a variety of developmental processes. The action of SINAT2 may explain the recalcitrance of AtRAP2.2 protein levels to change by altering AtRAP2.2 transcription.
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