Molecular Regulation Of RNA Metabolism

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications & trials

Abstract

Investigation of the RNA polymerase III (Pol III) system of all eukaryotes examined including human cells, the model studied in our lab, includes the RNA binding protein known as La protein. RNA polymerase III is solely responsible for synthesis of all cellular tRNAs as well as some essential noncoding (nc)RNAs. This research was was continued, focusing on transcription termination and its links to posttranscriptional 3' processing especially via the La protein which in humans is a prevelant target of autoantibodies in patients suffering from rheumatic disorders such as neonatal lupus, systemic lupus erythematosus and Sjogrens syndrome. RNAP III produces small RNAs, including the 5S rRNA and tRNAs essential for protein synthesis, as well as other cellular RNAs and certain virus-encoded transcripts. In addition to its conventional role in synthesis of essential tRAs and other small RNAs, a form of Pol III in the cytoplasm serves as an important sensor intermediary of innate immunity signalling. RNAP III is a complex enzyme composed of seventeen subunits with multiple catalytic activities. Human La antigen is a regulatory phosphoprotein that has been shown to serve to link 3' processing with termination by RNAP III, and directing the posttranscripional maturation pathway of the transcripts, the latter of which includes end-processing, intron removal, base modification, and proper RNA folding. The La phosphoprotein interacts with RNAP III transcripts by recognizing their 3 terminal UUU-OH motifs (which result from transcription termination), found at the ends of all newly synthesized pol III transcripts. Nonphosphorylated La is localized in the cytoplasm where it interacts with certain cellular and viral mRNAs including HIV RNA, hepatitis C RNA. poliovirus mRNA and others. La interacts with the internal ribosome entry sites (IRES) of viral and cellular mRNAs to modulate their translation. Some viral-encoded factors, including the adenovirus E1A protein, modulate pol III activity. Poliovirus protease-3 protein cleaves the phosphorylation site and nuclear localization signals away from the body of the La antigen, leading to a mainly cytoplasmic localization that facilitates La-mediated translation of poliovirus mRNA. We are interested in the tRNA anticodon modifications that impact the codon-usage specific translation of specific mRNAs involved in growth and development, with specific emphasis on TRIT1 the enzyme that adds isopentenyl group to A37 of the anticodon of a subset of tRNAs. In addition, we study another tRNA modification enzyme, Trm1 which creates dimethyl-guanosine at position 26 of a subset of tRNAs. Understanding the mechanisms by which La and other pol III subunits function in RNA production pathways, and how these pathways are controlled during normal development and cellular proliferation, are major goals of this Section. We extended our focus to La-related proteins (LARPs), including human LARP4 and LARP5, mRNA-associated proteins. LARP4 was shown to exhibit cellular activity to promote the net lengthening of poly(A) tails (PATs) on ribosome protein (rp) mRNAs and other mRNAs. Mouse cells genetically deleted of LARP4 exhibit shorter rpmRNA PATs and faster decay than their WT counterpart cells.