The general aims of this project are to identify the mechanisms of light activated translation of organelle mRNAs. Previous biochemical and genetic experiments have identified key components required for translation of the chloroplast psbA mRNA. Under this proposal we will continue the characterization of psbA translation with the ultimate goal of identifying the complete set of proteins and RNA elements required for translation of chloroplast mRNAs. We will also investigate the molecular interactions of these components that result in light activated translation. The completion of this set of experiments should provide the basis for a detailed molecular model of translation of organelle mRNAs. Understanding chloroplast translation may also prove to be a paradigm for understanding prokaryotic translation, and for understanding the molecular basis of redox regulation of gene expression. These studies should also impact our ability to exploit plant chloroplasts as a vehicle for the expression of recombinant proteins, that potentially could be used as therapeutic agents in human health. The sets of experiments to address chloroplast translation can be divided into five specific aims: 1. Identify the mechanism by which the chloroplast poly(A) binding protein activates psbA translation. 2. Isolate a complete set of psbA mRNA binding proteins, and clone the encoding genes. 3. Identify the RNA elements required for psbA translation, and characterize the mechanism by which these elements function. 4. Identify the roles of psbA associated proteins in translation by genetic analysis. 5. Characterize the role of RNA elements and trans-acting factors in translation initiation complex formation

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM054659-05A1
Application #
6370525
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Rhoades, Marcus M
Project Start
1997-05-01
Project End
2005-11-30
Budget Start
2001-12-01
Budget End
2002-11-30
Support Year
5
Fiscal Year
2002
Total Cost
$333,360
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Coragliotti, Anna T; Beligni, Maria Veronica; Franklin, Scott E et al. (2011) Molecular factors affecting the accumulation of recombinant proteins in the Chlamydomonas reinhardtii chloroplast. Mol Biotechnol 48:60-75
Rasala, Beth A; Muto, Machiko; Lee, Philip A et al. (2010) Production of therapeutic proteins in algae, analysis of expression of seven human proteins in the chloroplast of Chlamydomonas reinhardtii. Plant Biotechnol J 8:719-33
Hannon, Michael; Gimpel, Javier; Tran, Miller et al. (2010) Biofuels from algae: challenges and potential. Biofuels 1:763-784
Beligni, Maria Veronica; Mayfield, Stephen P (2008) Arabidopsis thaliana mutants reveal a role for CSP41a and CSP41b, two ribosome-associated endonucleases, in chloroplast ribosomal RNA metabolism. Plant Mol Biol 67:389-401
Mayfield, Stephen P; Manuell, Andrea L; Chen, Stephen et al. (2007) Chlamydomonas reinhardtii chloroplasts as protein factories. Curr Opin Biotechnol 18:126-33
Manuell, Andrea L; Beligni, Maria Veronica; Elder, John H et al. (2007) Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast. Plant Biotechnol J 5:402-12
Manuell, Andrea L; Quispe, Joel; Mayfield, Stephen P (2007) Structure of the chloroplast ribosome: novel domains for translation regulation. PLoS Biol 5:e209
Barnes, Dwight; Franklin, Scott; Schultz, Jason et al. (2005) Contribution of 5'- and 3'-untranslated regions of plastid mRNAs to the expression of Chlamydomonas reinhardtii chloroplast genes. Mol Genet Genomics 274:625-36
Somanchi, Aravind; Barnes, Dwight; Mayfield, Stephen P (2005) A nuclear gene of Chlamydomonas reinhardtii, Tba1, encodes a putative oxidoreductase required for translation of the chloroplast psbA mRNA. Plant J 42:341-52
Manuell, Andrea L; Yamaguchi, Kenichi; Haynes, Paul A et al. (2005) Composition and structure of the 80S ribosome from the green alga Chlamydomonas reinhardtii: 80S ribosomes are conserved in plants and animals. J Mol Biol 351:266-79

Showing the most recent 10 out of 23 publications