Proper regulation of histone biosynthesis during the cell cycle is critical for coordinating chromatin assembly and DNA replication. Overproduction or underproduction of histone protein results in replication stress and genome instability that contributes to the development of cancer. In this proposal, we undertake a comprehensive analysis of replication-dependent (RD) histone mRNA biosynthesis and how this process is regulated during the cell cycle in the context of animal development. RD-histone mRNAs are the only eukaryotic mRNAs in animal cells that lack a polyA tail, ending instead in a conserved stem loop structure. In contrast, mRNAs for histone variants such as H3.3 and H2A.Z are encoded by polyadenylated mRNAs that are not cell cycle regulated. Generation of the unique RD-histone mRNA 3? end results from the activity of an evolutionary conserved set of pre-RNA processing factors. The genes encoding all five RD-histone proteins are clustered in metazoan genomes, and transcription and pre- mRNA processing factors required for histone mRNA biosynthesis are organized into a nuclear body (the histone locus body or HLB) that assembles at these gene clusters. We will determine the requirements for the coordinate synthesis of the RD-histone mRNAs using both biochemical and genetic approaches in Drosophila, with a particular focus on the role that the HLB plays in histone transcription and pre-mRNA processing. Histone gene clusters provide a system in which one can readily study the expression of a tightly regulated set of genes at all levels of mRNA biosynthesis, from the organization of genes within the nucleus through activation of transcription, pre-mRNA processing and transcription termination. Our Drosophila experimental paradigm permits the in vivo interrogation of these fundamental processes in gene expression in ways that are unavailable in other experimental systems.

Public Health Relevance

Each time DNA is replicated during the process of cell proliferation, histone proteins are synthesized in large amounts to package newly made DNA into chromosomes. We will combine biochemical and genetic studies in the fruit fly Drosophila to study the mechanisms and regulation of the synthesis of histones proteins. Factors controlling histone synthesis are critical for stable inheritance of genetic information during organism development, and thus for the suppression of human disease. Because of high evolutionary conservation, our studies in flies will allow us to understand the function of these factors in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM058921-21
Application #
10130189
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
1999-05-01
Project End
2024-11-30
Budget Start
2020-12-09
Budget End
2021-11-30
Support Year
21
Fiscal Year
2021
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Biochemistry
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Meserve, Joy H; Duronio, Robert J (2018) Fate mapping during regeneration: Cells that undergo compensatory proliferation in damaged Drosophila eye imaginal discs differentiate into multiple retinal accessory cell types. Dev Biol 444:43-49
Skrajna, Aleksandra; Yang, Xiao-Cui; Dadlez, Michal et al. (2018) Protein composition of catalytically active U7-dependent processing complexes assembled on histone pre-mRNA containing biotin and a photo-cleavable linker. Nucleic Acids Res 46:4752-4770
Duronio, Robert J; Marzluff, William F (2017) Coordinating cell cycle-regulated histone gene expression through assembly and function of the Histone Locus Body. RNA Biol 14:726-738
Duronio, Robert J; O'Farrell, Patrick H; Sluder, Greenfield et al. (2017) Sophisticated lessons from simple organisms: appreciating the value of curiosity-driven research. Dis Model Mech 10:1381-1389
Rieder, Leila E; Koreski, Kaitlin P; Boltz, Kara A et al. (2017) Histone locus regulation by the Drosophila dosage compensation adaptor protein CLAMP. Genes Dev 31:1494-1508
Skrajna, Aleksandra; Yang, Xiao-Cui; Bucholc, Katarzyna et al. (2017) U7 snRNP is recruited to histone pre-mRNA in a FLASH-dependent manner by two separate regions of the stem-loop binding protein. RNA 23:938-951
Meserve, Joy H; Duronio, Robert J (2017) A population of G2-arrested cells are selected as sensory organ precursors for the interommatidial bristles of the Drosophila eye. Dev Biol 430:374-384
Marzluff, William F; Koreski, Kaitlin P (2017) Birth and Death of Histone mRNAs. Trends Genet 33:745-759
Tatomer, Deirdre C; Terzo, Esteban; Curry, Kaitlin P et al. (2016) Concentrating pre-mRNA processing factors in the histone locus body facilitates efficient histone mRNA biogenesis. J Cell Biol 213:557-70
Swanson, Christina I; Meserve, Joy H; McCarter, Patrick C et al. (2015) Expression of an S phase-stabilized version of the CDK inhibitor Dacapo can alter endoreplication. Development 142:4288-98

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