Collaborative Research

  • Functional Proteomics

    The wealth of sequenced genomes has resulted in the development of powerful genomic approaches to define the functions of the individual genes, to deduce the networks of interactions between genes and to mine additional information encoded in the genome. For example, a collection of 5,854 sequence-verified yeast expression plasmids opened the door to study signals within genes that regulate their expression, as well as to develop yeast as a host for expression of proteins for biochemical and structural analysis.

  • High Throughput Purification of Proteins for Biochemical and Structural Analysis

    Recent work in this lab and the lab of E. Grayhack has focused on establishing yeast as the eukaryote of choice for high throughput purification of yeast and other eukaryotic proteins for biochemical analysis and structural biology. Although expression in E. coli has been used for biochemical analysis and for the determination of a large number of protein structures, expression of eukaryotic proteins in E. coli often results in limited solubility, as well as in the absence of post-translational modifications. To establish yeast as an organism for high throughput cloning, expression and purification, we have developed several methods.

  • Mechanism of Protein Synthesis

    All ribosomes have a ribonucleoprotein structure made up of two unequal subunits, both of which contain three binding sites for tRNA: the A (aminoacyl), P (peptidyl) and E (exit) sites. Sequential movement of tRNAs from the A site to the P site to the E site is coupled with movement of their associated codons in the mRNA.

  • Post-transcriptional Gene Regulation by Double-stranded RNA-binding Proteins

    • Maquat Lab Normal & Disease-Associated RNA Decay
    • Kielkopf Lab Pre-mRNA Splicing for Treatment of Human Disease

    We have found that STAU1, which is a double-stranded RNA binding protein, recruits the NMD factor UPF1 to certain mRNA 3'-untranslated regions (3'UTRs) so as to elicit SMD in a translation-dependent mechanism (reviewed in Park and Maquat, 2013, Wiley Interdiscip Rev RNA 4:423-435). Using microarray analyses, we have identified a number of mRNAs that are naturally down-regulated by SMD.

  • The Biological Roles of tRNA Modifications

    tRNA is the most highly modified class of RNA species, and modifications are found in tRNAs from all organisms that have been examined. Yeast cytoplasmic tRNAs have 25 biochemically distinct modifications, and the average tRNA has 13 modified residues. These modifications are highly conserved in eukaryotes, including humans, and many of the corresponding genes have been identified in this and other labs. It is known from previous work in the field that several modifications in and around the anticodon loop have crucial roles in various aspects of translation, while several other modifications remote from the anticodon loop have specific roles in tRNA folding and/or stability. However, the precise roles of many other individual modifications are only poorly understood, and are under investigation in our lab.

Recent Publications