title: Reciprocal regulation of protein synthesis and carbon metabolism for thylakoid membrane biogenesis.
abstract: Metabolic control of gene expression coordinates the levels of specific gene products to meet cellular demand for their activities. This control can be exerted by metabolites acting as regulatory signals and/or a class of metabolic enzymes with dual functions as regulators of gene expression. However, little is known about how metabolic signals affect the balance between enzymatic and regulatory roles of these dual functional proteins. We previously described the RNA binding activity of a 63 kDa chloroplast protein from Chlamydomonas reinhardtii, which has been implicated in expression of the psbA mRNA, encoding the D1 protein of photosystem II. Here, we identify this factor as dihydrolipoamide acetyltransferase (DLA2), a subunit of the chloroplast pyruvate dehydrogenase complex (cpPDC), which is known to provide acetyl-CoA for fatty acid synthesis. Analyses of RNAi lines revealed that DLA2 is involved in the synthesis of both D1 and acetyl-CoA. Gel filtration analyses demonstrated an RNP complex containing DLA2 and the chloroplast psbA mRNA specifically in cells metabolizing acetate. An intrinsic RNA binding activity of DLA2 was confirmed by in vitro RNA binding assays. Results of fluorescence microscopy and subcellular fractionation experiments support a role of DLA2 in acetate-dependent localization of the psbA mRNA to a translation zone within the chloroplast. Reciprocally, the activity of the cpPDC was specifically affected by binding of psbA mRNA. Beyond that, in silico analysis and in vitro RNA binding studies using recombinant proteins support the possibility that RNA binding is an ancient feature of dihydrolipoamide acetyltransferases. Our results suggest a regulatory function of DLA2 in response to growth on reduced carbon energy sources. This raises the intriguing possibility that this regulation functions to coordinate the synthesis of lipids and proteins for the biogenesis of photosynthetic membranes.
authors: Molecular Plant Sciences, Ludwig-Maximilians-University Munich, Planegg-Martinsried, Germany.
authors: Alexandra-Viola
authors: AV
authors: Bohne
authors: Christian
authors: C
authors: Schwarz
authors: Marco
authors: M
authors: Schottkowski
authors: Michael
authors: M
authors: Lidschreiber
authors: Markus
authors: M
authors: Piotrowski
authors: William
authors: W
authors: Zerges
authors: Jörg
authors: J
authors: Nickelsen
mesh_terms: D002244:Carbon / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000378:metabolism; D008856:Microscopy, Fluorescence; D001678:Organelle Biogenesis; D045332:Photosystem II Protein Complex / Q000378:metabolism; D010940:Plant Proteins / Q000096:biosynthesis; D014176:Protein Biosynthesis; D020524:Thylakoids / Q000378:metabolism*
chemical_list: D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D002244:Carbon