Seminars | Biochemistry

RNA research at Madison thrives in many labs. RNA MaxiGroup is intended to bring those labs together to exchange ideas and methods. Presentations are without slides to encourage discussion.

Anyone interested is welcome, even if they are not particularly interested in RNA in general - graduate students, post-docs, staff and faculty.

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Light snacks are provided before seminar, beginning at 5:15 pm

Organizers - Marv Wickens (Chair), Dave Brow, Sam Butcher, Jim Dahlberg, Aaron Hoskins, Judith Kimble, Andrew Mehle and Mike Sheets.

RNA-protein interactions inside a cell identified by “RNA tagging” A chimeric protein consisting of an RNA-binding protein (RBP) fused to a U-adding enzyme (PUP) is expressed in a cell. Binding of that RBP-PUP protein to an mRNA via a binding element (blue rectangle) marks that RNA with U’s. From the Marv Wickens lab

Control of many mitochondrial functions through the binding of a single PUF protein to many mRNAs. A multi-omic study integrated RNA tagging, CLIP, proteomics, lipidomics and metabolomics to identify mRNAs controlled by the PUF protein, Puf3p. These include the protein import machinery, the mitochondrial ribosome, and oxidative phosphorylation. Collaboration between Wickens and Pagliarini labs

RNA regulation drives cell fate decisions in the germ cells in all animals. This general principle emerged from work on the nematode germline, which is pictured here with its stem cells at one end (blue) and gametes at the other (oocytes, green; sperm, red). From the Judith Kimble lab

Image of U1 snRNP and branchpoint bridging protein

U1 snRNP (red) and branchpoint bridging protein (green) binding events on 30 single molecules of pre-mRNA, seen by Colocalization Single Molecule Spectroscopy. Spliceosome E complex formation is noted by the blue bars. From the Aaron Hoskins lab

Investigating the function of RNA-binding proteins in neurons. From the Jill Wildonger lab

Model of the yeast Sen1-dependent termination complex scanning a nascent RNA polymerase II (Pol II) transcript. Binding of terminator elements in the transcript to Nrd1 and Nab3 elicits termination of transcription in a manner dependent on the helicase activity of Sen1 and glutamate 108 of the Rpb11 subunit. From the Dave Brow lab

Protein curvature in a common architecture determines the specificity of RNA-protein interactions. Biochemical, genetic and structural analyses of multiple PUF proteins bound to their RNA targets reveals that the curvature of the proteins is critical in determining which RNAs each protein binds. Wickens and T. Hall labs

Human cardiomyocyte derived from IPS cells. From the Mike Sheets lab

Image of new families of nucleotidyl transferase enzymes

New families of nucleotidyl transferase enzymes. The enzyme GLD-2 was discovered in C. elegans through a combination of genetics and biochemistry (collaboration of Kimble and Wickens labs). This led to tethered function assays that revealed families of regulatory enzymes that add A’s (PAPs) or U’s (PUPs) to a wide range of RNAs. Both families have widespread roles in biology, including development, the nervous system, and disease. From the Wickens lab

Tracks of machinery that support local translation in dendrites. From the Jill Wildonger lab

The Hoskins Lab studies hotspot mutations in the spliceosome protein SF3b1 that cause myelodysplastic syndromes (MDS) and other blood cancers

Aaron Hoskins (left) and Josh Larson (right) use their multi-wavelength, micro-mirror CoSMoS microscope for single molecule studies of spliceosome assembly

Structure of 5’ UMP bound in the active site of Usb1. From the Sam Butcher lab

Crystal structure of the yeast Prp24-U6 RNA complex (PDB ID: 4n0t) illustrating the topological entanglement of RNA and protein. From the Dave Brow lab

Fluorescent influenza A reporter viruses were designed to enable real-time detection of co-infected cells. Live cell imaging identifies cells infected by both a GFP and RFP reporter viruses. From the Andrew Mehle Lab

Structure of U6 RNA bound to Prp24 from the Sam Butcher lab

Image of rewiring of RNA-protein networks

Evolutionary rewiring of RNA-protein networks. Direct biochemical analyses of RNA-protein interactions in two highly diverged fungal species demonstrates that a biological function controlled by Puf3p in one species is controlled by a different protein, Puf5p, in the other. Collaboration between the Wickens and Gasch labs

Photo of Bicc1 particles in Retinal pigment epithelial cells. From the Mike Sheets lab

Cytoplasmic particles formed by the human Bicaudal-C translational repressor protein

The Andrew Mehle Lab “hacked” the influenza virus genome to encode the very small and bright NanoLuciferase. In the example shown here, reporter virus was imaged following infection in an embyronated chicken eggs. Blood vessels and the embryo at the bottom are clearly visible. Artistic license was used to depict virus streaking away from the open top of the infected egg

The Hoskins Lab studies how the spliceosome protein SF3b1 recognizes the RNA duplex formed between the intronic branch site and the U2 snRNA

Figure of single molecule colocalization

Data from the Hoskins Lab depicting single molecule colocalization of the yeast U1 snRNP and Branchpoint Bridging Protein (BBP) during spliceosome E complex formation