Many bacteria have a sophisticated mechanism for metabolic regulation that involves microcompartments. These bacterial microcompartments are made up of unique protein shells similar in structure to a viral capsid and enclose two or more enzymes in a particular reaction pathway. Carboxysomes from cyanobacteria and chemoauxotrophs are the most well known bacterial microcompartments. Within the last few years, the crystal structures of several shell components have been solved and have led to remarkable progress in the knowledge of subunit assembly and function of individual components. So far, there has been no report of heterologous assemblies of bacterial microcompartments in eukaryotic organisms. This project will target multiple carboxysome proteins into chloroplasts. This study will employ transient expression of foreign proteins fused with fluorescent tags by agroinfiltration technique. Any molecular complex assembled from the expressed proteins will be monitored with confocal microscopy, and the structures formed will be characterized by electron microscopy. In addition, this work will generate stable transgenic lines, in which carboxysome proteins are expressed from either the nucleus or the chloroplast genome. By expressing all necessary carboxysome structural components, this project will attempt to assemble the carboxysome microcompartment shells in eukaryotes. In addition, this study will determine whether it is possible to target foreign proteins to the carboxysome shells. The proposed study is an important step for engineering novel microcompartments and will have potential future applications in areas such as drug delivery, metabolic engineering and biotechnology.

Public Health Relevance

Many bacteria have a sophisticated mechanism for metabolic regulation that involves microcompartments made up of unique protein shells similar in structure to a viral capsid. The goal of this project is to engineer the structural components of a bacterial microcompartment known as carboxysome so that it can be introduced into other organisms. The proposed study will have possible future applications in areas such as drug delivery, metabolic engineering and biotechnology.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM103019-02
Application #
8686611
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Sakalian, Michael
Project Start
2013-07-01
Project End
2015-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Cornell University
Department
Biochemistry
Type
Earth Sciences/Resources
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Occhialini, Alessandro; Lin, Myat T; Andralojc, P John et al. (2016) Transgenic tobacco plants with improved cyanobacterial Rubisco expression but no extra assembly factors grow at near wild-type rates if provided with elevated CO2. Plant J 85:148-60
Hanson, Maureen R; Lin, Myat T; Carmo-Silva, A Elizabete et al. (2016) Towards engineering carboxysomes into C3 plants. Plant J 87:38-50
Lin, Myat T; Occhialini, Alessandro; Andralojc, P John et al. (2014) ?-Carboxysomal proteins assemble into highly organized structures in Nicotiana chloroplasts. Plant J 79:1-12
Lin, Myat T; Occhialini, Alessandro; Andralojc, P John et al. (2014) A faster Rubisco with potential to increase photosynthesis in crops. Nature 513:547-50