Unlocking the Mechanism of tRNA Translocation Through the Ribosome Using Large-Scale Molecular Simulation
Karissa Sanbonmatsu, NMC Affiliated Research Scientist, LANL Staff Scientist
The ribosome is one of the largest and highly conserved molecular machines, whose function is essential to all life. Its operational principles, however, remain largely a mystery. Among the myriad of steps catalyzed by the ribosome during protein synthesis, translocation is considered the ‘holy grail’ by many. Translocation is an intrinsic property of the ribosome, involving complex conformational changes of the two ribosome subunits that move the mRNA and tRNA substrates by precisely three nucleotides in a unidirectional fashion. Recently, studies demonstrated that the majority of translocation occurs during head swivel (intra-subunit rotation within the small subunit). Understanding the connection between head swivel and mRNA/tRNA movement will unlock a key operational principle of translocation. While there is growing consensus that head swivel is crucial to translocation, a direct relationship between these conformational rearrangements and the translocation reaction coordinate is lacking. We will use closely integrated molecular simulation and single-molecule FRET imaging studies to bridge this gap. Development of Attenuated HCMV/HIV Vectors for an HIV/AIDS Vaccine
Discriminative Biomarker Assays for Detection of Shiga toxin-producing Escherichia coli
A Rapid Nucleic Acid-based Assay for the Detection and Characterization of E. Coli.
Shiga toxin-producing E. coli (STEC) are a serious threat to our food supply and public health, causing more than 265,000 infections each year. NMC participates with UNM on a large USDA grant that seeks to understand how E. coli pathogens travel throughout the beef production process and how outbreaks occur. The program seeks to find ways to prevent illness and improve the safety of our nation's food supply. The NMC/UNM team is applying LANL developed technology to develop and implement rapid detection technologies for pre-harvest, post-harvest and consumer environments. The team has developed a rapid nucleic acid-based assay for the detection and characterization of STEC in the beef chain. This assay is the multiplex oligonucleotide ligation-PCR (MOL-PCR) and unlike other ligation-based assays that require multiple steps, MOL-PCR consists of a single tube reaction.
Development of Novel Vector / Protein HIV-1 Vaccines
This program develops novel vector/protein HIV-1 vaccines by combining the vaccine platform technologies of Beth Israel Deaconess Medical Center (BIDMC) and GlaxoSmithKline Biologicals (GSK). The combination of BIDMC and GSK vectors, proteins, adjuvants, and expertise will result in a new academic-industry partnership that will rapidly translate innovative HIV-1 vaccine product concepts into clinical trials. The goal of this program is to define an optimized vector/protein HIV-1 Env vaccine with immediate clinical development potential. The project hypothesizes that priming with a novel adenovirus (Ad) vector and boosting with a stable Env trimer optimized for immunologic coverage of global virus diversity and formulated with the GSK AS01B Adjuvant System will induce potent and protective Env-specific antibody responses. The Ad vector priming will prove superior to poxvirus vector priming for a subsequent Env protein boost. This research will optimize each component of this vaccine regimen systematically in a logical, focused product development program involving a series of nonhuman primate challenge studies.
A High Throughput Pipeline to Select Renewable Recombinant Polyclonal Antibodies
This joint LANL/NMC research program develops high throughput affinity reagent selection and screening capabilities. In this project, the team is creating new methods to select and screen for antibodies. The research creates new antibody libraries, develops new selection methods using flow cytometry and explores novel ways to use these antibodies.
From the End to the Beginning: Termination and Ribosome Recycling in Eukaryotic Translation
Protein synthesis research is important to understanding the origin of many human diseases and devising treatments for them. This project examines a poorly understood step in protein synthesis: the structure of intermediate states in termination and eukaryote ribosome recycling. The research will model the transition between these intermediate states.