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 #
5R01CA139322-06
Application #
8464022
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Hildesheim, Jeffrey
Project Start
2009-06-01
Project End
2014-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
6
Fiscal Year
2013
Total Cost
$275,440
Indirect Cost
$100,465
Name
University of Vermont & St Agric College
Department
Biochemistry
Type
Schools of Medicine
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
05405
Kapinas, Kristina; Kim, Heesun; Mandeville, Matthew et al. (2015) microRNA-mediated survivin control of pluripotency. J Cell Physiol 230:63-70
Stumpff, Jason; Ghule, Prachi N; Shimamura, Akiko et al. (2014) Spindle microtubule dysfunction and cancer predisposition. J Cell Physiol 229:1881-3
Ghule, Prachi N; Xie, Rong-Lin; Medina, Ricardo et al. (2014) Fidelity of histone gene regulation is obligatory for genome replication and stability. Mol Cell Biol 34:2650-9
Dudakovic, Amel; Camilleri, Emily; Riester, Scott M et al. (2014) High-resolution molecular validation of self-renewal and spontaneous differentiation in clinical-grade adipose-tissue derived human mesenchymal stem cells. J Cell Biochem 115:1816-28
Kapinas, Kristina; Grandy, Rodrigo; Ghule, Prachi et al. (2013) The abbreviated pluripotent cell cycle. J Cell Physiol 228:9-20
Lian, Jane B; Stein, Gary S; van Wijnen, Andre J et al. (2012) MicroRNA control of bone formation and homeostasis. Nat Rev Endocrinol 8:212-27
Stein, Gary S; Stein, Janet L; van J Wijnen, Andre et al. (2012) The architectural organization of human stem cell cycle regulatory machinery. Curr Pharm Des 18:1679-85
Liu, Li-Jun; Xie, Ronglin; Hussain, Sadiq et al. (2011) Functional coupling of transcription factor HiNF-P and histone H4 gene expression during pre- and post-natal mouse development. Gene 483:1-10
Stein, Gary S; Stein, Janet L; van Wijnen, Andre J et al. (2011) An architectural genetic and epigenetic perspective. Integr Biol (Camb) 3:297-303
Ghule, Prachi N; Medina, Ricardo; Lengner, Christopher J et al. (2011) Reprogramming the pluripotent cell cycle: restoration of an abbreviated G1 phase in human induced pluripotent stem (iPS) cells. J Cell Physiol 226:1149-56

Showing the most recent 10 out of 12 publications