Entry into S phase is a key event for regulation of mammalian cell proliferation. The controlled activation of histone gene expression at the G1/S phase transition is essential for chromatin packaging of nascent DNA. Our program has identified HiNF-P as the principal transcriptional regulator of histone genes, the end-point molecule for the cyclin E/ CDK2/ p220NPAT pathway, and HiNF-P deficiency impairs cell cycle progression through S phase. The discovery and characterization of HiNF-P represents one of the most important accomplishments of our program and the proposed studies will build on this major result to open a new dimension to cell cycle control. Our central hypothesis is that the cyclin E/ CDK2/ p220NPAT/ HiNF-P gene regulatory cascade is a principal cell cycle pathway that functions to achieve competency for histone H4 biosynthesis to support packaging of DNA as chromatin. To address this hypothesis, we will combine biochemical, molecular, cellular and in vivo genetic strategies to define how the HiNF-P/p220NPAT pathway supports S phase activation and progression. First, we will characterize protein-protein interaction domains and post-translational modifications that control the activity of the HiNF-P/p220NPAT complex, as well as the in situ integration of regulatory signals at subnuclear sites ('Histone Locus Bodies') that mediate histone gene transcription and mRNA processing (Specific Aim 1). We will then examine how HiNF-P deficiency affects cell cycle progression in normal and tumor cells and address HiNF-P related mechanisms involved in regulating cell proliferation (Specific Aim 2). To establish the in vivo relevance of the HiNF-P pathway, we will investigate whether HiNF-P is important for normal development in vivo during embryogenesis and post-natal growth (Specific Aim 3). The three proposed aims are designed to provide an integrated understanding of the molecular, cellular, and biological contributions of the cyclin E/CDK2/p220NPAT/HiNF-P pathway to cell proliferation in normal and tumor cells.

Public Health Relevance

Relevance: Cell proliferation is regulated by a complex and interdependent series of biochemical events involving cell cycle-stage specific modifications in gene expression. The S-phase specific expression of histone genes is both temporally and functionally coupled with DNA replication. Cell cycle dependent modulation of histone gene transcription via HiNF-P and p220NPAT provides the initial rate-limiting step in the induction of histone protein synthesis at the G1/S phase transition. The HiNF-P dependent mechanisms we are investigating represent fundamentally novel pathways involved in cell cycle control. Our studies will provide insight into mechanisms that control competency for S phase progression and may yield novel targets for selective treatment of diseases, particularly cancer, in which growth control is compromised.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA139322-04
Application #
8247176
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Hildesheim, Jeffrey
Project Start
2009-06-01
Project End
2012-07-31
Budget Start
2012-05-01
Budget End
2012-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$127,982
Indirect Cost
$60,144
Name
University of Massachusetts Medical School Worcester
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Zaidi, Sayyed K; Boyd, Joseph R; Grandy, Rodrigo A et al. (2016) Expression of Ribosomal RNA and Protein Genes in Human Embryonic Stem Cells Is Associated With the Activating H3K4me3 Histone Mark. J Cell Physiol 231:2007-13
VanOudenhove, Jennifer J; Medina, Ricardo; Ghule, Prachi N et al. (2016) Transient RUNX1 Expression during Early Mesendodermal Differentiation of hESCs Promotes Epithelial to Mesenchymal Transition through TGFB2 Signaling. Stem Cell Reports 7:884-896
VanOudenhove, Jennifer J; Grandy, Rodrigo A; Ghule, Prachi N et al. (2016) Unique Regulatory Mechanisms for the Human Embryonic Stem Cell Cycle. J Cell Physiol :
Barutcu, A Rasim; Fritz, Andrew J; Zaidi, Sayyed K et al. (2016) C-ing the Genome: A Compendium of Chromosome Conformation Capture Methods to Study Higher-Order Chromatin Organization. J Cell Physiol 231:31-5
Ghule, Prachi N; Xie, Rong-Lin; Colby, Jennifer L et al. (2016) Maternal expression and early induction of histone gene transcription factor Hinfp sustains development in pre-implantation embryos. Dev Biol 419:311-320
Grandy, Rodrigo A; Whitfield, Troy W; Wu, Hai et al. (2016) Genome-Wide Studies Reveal that H3K4me3 Modification in Bivalent Genes Is Dynamically Regulated during the Pluripotent Cell Cycle and Stabilized upon Differentiation. Mol Cell Biol 36:615-27
Van Oudenhove, Jennifer J; Grandy, Rodrigo A; Ghule, Prachi N et al. (2016) Lineage-Specific Early Differentiation of Human Embryonic Stem Cells Requires a G2 Cell Cycle Pause. Stem Cells 34:1765-75
Scott, Robert E; Ghule, Prachi N; Stein, Janet L et al. (2015) Cell cycle gene expression networks discovered using systems biology: Significance in carcinogenesis. J Cell Physiol 230:2533-42
Ghule, Prachi N; Xie, Rong-Lin; Colby, Jennifer L et al. (2015) p53 checkpoint ablation exacerbates the phenotype of Hinfp dependent histone H4 deficiency. Cell Cycle 14:2501-8
Kapinas, Kristina; Kim, Heesun; Mandeville, Matthew et al. (2015) microRNA-mediated survivin control of pluripotency. J Cell Physiol 230:63-70

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