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Principal Investigator: Bruce Alan Knutson
Organization: UPSTATE MEDICAL UNIVERSITY
Fiscal Year: 2024
Award: $122,450
Funding agency: National Institute of General Medical Sciences
Project Summary
Eukaryotic RNA polymerase I (Pol I) transcribes ribosomal RNA, a key component of ribosomes. Pol I transcription
accounts for the majority of the total RNA in cells, and its upregulation in human cells is a hallmark of cancer while
its downregulation is a hallmark of several developmental disorders. Pol I transcription is understudied compared
to transcription by Pol II and even Pol Ill. Our preliminary work suggests fundamental differences between Pol I
and Pols II and 111 that are the basis for this proposal. Our broad long-term objectives are to determine the molecular
mechanism of Pol I transcription and how its dysregulation leads to cancer and developmental disorders. There
are major gaps in our understanding of (1} the structural organization and architecture of Pol I transcription
complexes; (2) the mechanism for how Pol I initiation factors interact with rDNA, which encodes ribosomal RNA;
and (3) the molecular function of several key Pol I transcription factors in the activation process. The first rationale
for this work is that determining the mechanism and regulation of Pol I transcription will form the molecular basis
for understanding how Pol I defects lead to human disease. Our central hypothesis is that Pol I factors use a
unique mechanism to carry out transcription and their structure and function is different from the mechanisms
governing Pol II and Ill transcription. The second rationale is that understanding the Pol I transcription mechanism
at the most basic and fundamental levels will translate to a better understanding of the connection between Pol I
and cancer, leading to new cancer therapeutic strategies. Our proposed research will use a conceptually and
technically innovative cross-organismal and interdisciplinary approach that employs a combination of bioinformatic,
computational, molecular, biochemical, genetic, genomic, proteomic, and structural methods in the yeast and
human cells. Guided by strong preliminary studies, we will test two specific aims: (1) Determine the unique
"coactivator" role of TAT A-binding protein (TSP) in Pol I transcription, and (2) Determine the mechanism of Pol I
transcription activation. To accomplish these aims, we will use well-established and complementary approaches
to identify and map novel Pol I interactions in their native context. We will complement these studies with structural
modeling in combination with molecular, genetic, and biochemical functional assays to identify Pol I factor functions
conserved from yeast to humans. The proposed research is significant because it will lead to a detailed description
of the Pol I transcription mechanism and will provide a conceptual framework for understanding the link between
Pol I and human disease. Ultimately, this work will illuminate new avenues for diagnosis, potential interventions,
and the development of therapies targeting these novel protein-protein and protein-DNA interactions.
Terms: <Architecture><Assay><Basal Transcription Factor><Basal transcription factor genes><Binding Proteins><Bio-Informatics><Bioassay><Biochemical><Bioinformatics><Biological Assay><Blood Coagulation Factor I><Blood Coagulation Factor One><Blood Factor One><Cancer Cause><Cancer Etiology><Cancers><Cell Body><Cells><Coagulation Factor I><Coagulation Factor One><Complement><Complement Proteins><Complex><DNA Polymerase I><DNA Polymerase II><DNA Polymerase alpha><DNA Polymerase epsilon><DNA-Dependent DNA Polymerase I><DNA-Dependent DNA Polymerase II><DNA-Dependent RNA Polymerase I><DNA-Protein Interaction><Defect><Diagnosis><Disease><Disorder><Down-Regulation><Engineering / Architecture><Factor I><Factor One><Fibrinogen><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genetic><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic Transcription><Genetic defect><Genomics><Goals><Health><Human><Initiation Factors><Intervention><Intervention Strategies><Klenow Fragment><Knowledge><Lead><Ligand Binding Protein><Ligand Binding Protein Gene><Link><Malignant Neoplasms><Malignant Tumor><Maps><Methods><Modern Man><Molecular><Mutation><Non-Polyadenylated RNA><Pb element><Peptide Initiation Factors><Pol I><Pol II><Process><Protein Binding><Proteins><Proteomics><RNA><RNA Expression><RNA Gene Products><RNA Polymerase A><RNA Polymerase I><Regulation><Research><Ribonucleic Acid><Ribosomal DNA><Ribosomal RNA><Ribosomes><Role><Structural Models><Structure><Testing><Therapeutic><Transcription><Transcription Activation><Transcription Factor Proto-Oncogene><Transcription factor genes><Transcriptional Activation><Translating><Translation Initiation Factor><Translational Initiation Factor><Up-Regulation><Upregulation><Work><Yeasts><bound protein><complementation><develop therapy><developmental disease><developmental disorder><genetic condition><genetic disorder><genome mutation><heavy metal Pb><heavy metal lead><human disease><innovate><innovation><innovative><interdisciplinary approach><intervention development><interventional strategy><malignancy><multidisciplinary approach><neoplasm/cancer><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><rDNA><rRNA><social role><therapy development><transcription factor><treatment development>