title: Structure-function analysis of Rny1 in tRNA cleavage and growth inhibition.
abstract: T2 ribonucleases are conserved nucleases that affect a variety of processes in eukaryotic cells including the regulation of self-incompatibility by S-RNases in plants, modulation of host immune cell responses by viral and schistosome T2 enzymes, and neurological development and tumor progression in humans. These roles for RNaseT2's can be due to catalytic or catalytic-independent functions of the molecule. Despite this broad importance, the features of RNaseT2 proteins that modulate catalytic and catalytic-independent functions are poorly understood. Herein, we analyze the features of Rny1 in Saccharomyces cerevisiae to determine the requirements for cleaving tRNA in vivo and for inhibiting cellular growth in a catalytic-independent manner. We demonstrate that catalytic-independent inhibition of growth is a combinatorial property of the protein and is affected by a fungal-specific C-terminal extension, the conserved catalytic core, and the presence of a signal peptide. Catalytic functions of Rny1 are independent of the C-terminal extension, are affected by many mutations in the catalytic core, and also require a signal peptide. Biochemical flotation assays reveal that in rny1Δ cells, some tRNA molecules associate with membranes suggesting that cleavage of tRNAs by Rny1 can involve either tRNA association with, or uptake into, membrane compartments.
authors: Cancer Biology Graduate Interdisciplinary Program, University of Arizona, Tucson, Arizona, USA.
authors: Natalie
authors: N
authors: Luhtala
authors: Roy
authors: R
authors: Parker
mesh_terms: D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D009154:Mutation; D017433:Protein Structure, Secondary; D012343:RNA, Transfer / Q000378:metabolism*; D012260:Ribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D012260:Ribonucleases; C540806:Rny1 protein, S cerevisiae