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Title: Reciprocal Regulation of Protein Synthesis and Carbon Metabolism for Thylakoid Membrane Biogenesis

Journal Article · · PLoS Biology (Online)
 [1];  [1];  [2];  [1];  [3];  [2];  [1]
  1. Ludwig Maximilian University of Munich, Munich (Germany)
  2. Concordia University, Montreal (Canada)
  3. Ruhr University, Bochum (Germany)
+ Show Author Affiliations

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.

Research Organization:
USDOE Joint Genome Institute (JGI), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC); German Research Foundation (DFG); Natural Sciences and Engineering Council of Canada
Grant/Contract Number:
AC02-05CH11231; Ni390/4-2; 217566-03)
OSTI ID:
1904124
Journal Information:
PLoS Biology (Online), Vol. 11, Issue 2; ISSN 1545-7885
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (8)

Efficient expression of nuclear transgenes in the green alga Chlamydomonas: synthesis of an HIV antigen and development of a new selectable marker journal January 2016
Biogenesis of thylakoid membranes journal September 2015
Thylakoid biogenesis has joined the new era of bacterial cell biology journal January 2013
Complex(iti)es of the ubiquitous RNA-binding CSP41 proteins journal June 2014
Identification and Roles of Photosystem II Assembly, Stability, and Repair Factors in Arabidopsis journal February 2016
Chloroplast Genome of the Folk Medicine and Vegetable Plant Talinum paniculatum (Jacq.) Gaertn.: Gene Organization, Comparative and Phylogenetic Analysis journal April 2018
Deciphering the Proteotoxic Stress Responses Triggered by the Perturbed Thylakoid Proteostasis in Arabidopsis journal March 2021
The Chloroplast Ribonucleoprotein CP33B Quantitatively Binds the psbA mRNA journal March 2020

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
chloroplasts
RNA-binding proteins
Chlamydomonas reinhardtii
messenger RNA
enzyme metabolism
protein synthesis
gene expression
protein metabolism