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Principal Investigator: Yuan He
Organization: NORTHWESTERN UNIVERSITY
Fiscal Year: 2024
Award: $71,061
Funding agency: National Institute of General Medical Sciences
Summary
Transcription is the first step to read out the genetic information stored in the chromosome. In eukaryotes,
three multi-subunit RNA polymerases (Pols), Pol I, II, and III transcribe the 25S ribosomal RNA (rRNA),
protein-coding, and short non-coding RNAs such as transfer RNA (tRNA) and 25S rRNA genes, respectively. A
wealth of genetic and biochemical data accumulated over 50 years have uncovered most of the molecular
players involved and details of the intricate regulatory circuits. The recruitment and assembly of the initiation
complex at the promoter represent one of the key regulatory steps during gene expression. The Pol II initiation
machinery includes the activator-bound Mediator along with a series of general transcription factors (GTFs)
(TFIID, TFIIA, TFIIB, TFIIE, TFIIF, and TFIIH) that assemble into a ~4-megadalton (MDa) pre-initiation
complex (PIC) on core promoter DNA. On the other hand, the Pol III enzyme is recruited onto its promoter with
the help of corresponding GTFs (TFIIIC, TBP, BRF1, and B''). This proposal aims to investigate the
mechanism of transcription regulation by directly visualizing these transcriptional competent initiation
complexes using single-particle cryo-EM. Comparable pictures of these different but evolutionarily related
systems at this critical step of gene regulation will provide an unprecedented, comprehensive view of this key
process in the central dogma of molecular biology.
Terms: <25S rRNA><5S Gene-Specific Transcription Factor><5S RNA Gene Transcription Factor><90-kD Subunit Transcription Factor IIIB><ATP Hydrolysis><Address><Architecture><B-Related Factor 1><B-TFIID><BA2R><BEN gene><BEN protein><BRF Gene><BRF1><BRF1 Homolog><BRF1 Homolog Subunit of RNA Polymerase III Transcription Initiation Factor IIIB><BRF1 gene><BTF2 transcription factor><Basal Transcription Factor><Basal transcription factor genes><Binding><Biochemical><Biogenesis><CCG1><Cell Body><Cell Communication and Signaling><Cell Cycle Gene 1><Cell Nucleus><Cell Signaling><Cells><Cellular Expansion><Cellular Growth><Chemicals><Chromosomes><Code><Coding System><Complex><Core Assembly><Cryo-electron Microscopy><Cryoelectron Microscopy><DNA><DNA Molecular Biology><DNA-Dependent RNA Polymerase II><DNA-Dependent RNA Polymerases><DNA-Directed RNA Polymerase><Data><Deoxyribonucleic Acid><Electron Cryomicroscopy><Elements><Endogenous Interferon Beta><Engineering / Architecture><Enhancers><Enzyme Gene><Enzymes><Eukaryota><Eukaryote><Fibroblast Interferon><Formerly TAF3C><Functional RNA><GTF2B><GTF2B gene><GTF2I Repeat Domain-Containing Protein 1><GTF2IRD1><GTF2IRD1 gene><GTF3><GTF3A Protein><GTF3B><Gene Action Regulation><Gene Expression><Gene Expression Regulation><Gene Proteins><Gene Regulation><Gene Regulation Process><Gene Transcription><General Transcription Factor Gene><General Transcription Factor II-I Repeat Domain-Containing Protein 1><General Transcription Factor IIA><General Transcription Factor IIB Gene><General Transcription Factor III><General Transcription Factor IIIB 90kD Subunit><General Transcription Factor-IIB><General Transcription Factors><Genes><Genetic><Genetic Transcription><Goals><Human><IFN-Beta><IFN-β><IFNb><In Vitro><Interferon-beta><Interferon-β><Intracellular Communication and Signaling><MUSTRD1><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediator><Modern Man><Molecular><Molecular Biology><Molecular Interaction><Muscle TFII-I Repeat Domain-Containing Protein 1><Natural Interferon Beta><Natural human interferon beta><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nucleus><Organism><Origin of Life><Polymerase><Process><Production><Protein Biosynthesis><Protein Gene Products><Proteins><RNA><RNA Expression><RNA Gene Products><RNA Polymerase B><RNA Polymerase II><RNA Polymerase II Transcription Factor IIB Gene><RNA Polymerase II Transcription Factor-IIB><RNA Polymerases><RNA chemical synthesis><RNA synthesis><RNA, Ribosomal, 5S><RNA, ribosomal, 25S><Recycling><Resolution><Ribonucleic Acid><Ribosomal Peptide Biosynthesis><Ribosomal Protein Biosynthesis><Ribosomal Protein Synthesis><Ribosomal RNA><Ribosomal RNA Genes><Series><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Structure><System><TAF1><TAF1 RNA Polymerase II TATA Box Binding Protein-Associated Factor 250-kD><TAF1 gene><TAF2A><TAF3B2><TAF3B2 Gene><TAF3C><TAFII250><TAFIII90><TATA Box Binding Protein -Associated Factor 3C><TATA Box Binding Protein -Associated Factor RNA Polymerase III Subunit 2><TATA Box-Binding Protein-Associated Factor 2A><TATA Box-Binding Protein-Associated Factor, RNA Polymerase III, Subunit 2><TBP-Associated Factor RNA Polymerase II 250-kD><TBP-Associated Factor RNA Polymerase III 90kD><TBP-Associated Factor, RNA Polymerase III, 90-kD><TF2B Gene><TF3B90><TFIIA><TFIIB><TFIIB Gene><TFIID><TFIIE><TFIIF><TFIIH><TFIIIA><TFIIIB><TFIIIB90 Gene><TFIIIC><Techniques><Time><Transcription><Transcription Activation><Transcription Factor A RNA Polymerase II><Transcription Factor IIA><Transcription Factor IIB Gene><Transcription Factor IIIA><Transcription Factor IIIB><Transcription Factor Proto-Oncogene><Transcription Factor TFIIA><Transcription Factor TFIIB><Transcription Factor TFIIIA><Transcription Factor TFIIIB><Transcription Factor-IIB><Transcription Initiation><Transcription Initiation Factor IIA><Transcription Initiation Factor IIIB><Transcription Initiation Site><Transcription Regulation><Transcription Start Site><Transcription factor genes><Transcriptional Activation><Transcriptional Control><Transcriptional Regulation><Transfer RNA><Triplet Codon-Amino Acid Adaptor><Untranslated RNA><Visualization><WBSCR11><Yeasts><basic transcription factor 2><binding Factor For Early Enhancer><biological signal transduction><cell growth><crosslink><cryo-EM><cryoEM><cryogenic electron microscopy><genetic information><hBRF><human disease><interdisciplinary approach><living system><multidisciplinary approach><noncoding><novel><particle><polypeptide><promoter><promotor><protein synthesis><rRNA><rRNA Genes><reconstitute><reconstitution><recruit><resolutions><tRNA><transcription factor><transcription factor IIE><transcription factor IIH><transcription factor IIIC><transcription factor TFIIE><transcription factor TFIIF><transcription factor TFIIH><transcription factor TFIIIC><transfer Ribonucleic acids><translocase>