Engineering a fully-integrated translation system for production of proteins and peptides

Ribosomes are molecular machines. They produce cellular proteins. Like a computer that can run different programs, a ribosome can make different proteins encoded in the genetic message. The normal ribosome is generally not well-equipped for making specific designer proteins. The objective of this project is to investigate one approach to creating more effective designer protein production. This will involve the construction of a ribosome that is hard-wired for making one specific protein. This will be achieved by incorporating the genetic message into the ribosome itself. The design of the resulting ribosome could then be optimized to efficiently produce the desired protein. Undergraduates and high school students will participate in summer-long research projects supporting this award. In the cell, ribosomes associate with different mRNAs to synthesize different proteins required for cell growth and proliferation. Expression of designer proteins that do not contribute essential functions for cell metabolism acts to divert ribosomes away from producing those essential proteins. Diverting ribosomes in that way can reduce the fitness of the cell to grow and multiply. The strategy being tested is functional isolation of such ribosomes from ‘normal’ cellular translation. The isolation will be achieved by covalently attaching mRNA to ribosomal RNA. The resulting ribosome-mRNA hybrid (iRibo-T) is expected to translate only one specific mRNA and, therefore, is suitable for altering its catalytic capacity and other properties. The new design will be employed for optimizing expression of medically and industrially relevant poly-lysine containing proteins whose translation are problematic for the unaltered bacterial ribosomes. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.