The goal of this project is to understand the coordination of histone mRNA stability with DNA replication during the m ammalian cell cycle. The experiments included in this proposal are designed to: a) identify and delineate the specific regions of cell cycle-regulated histone messenger RNAs responsible for coupling metabolic stability with DNA synthesis, and b) characterize the molecules and mechanisms involved in the rapid degradation of histone mRNA that occurs in the absence of DNA replication. Fusion genes will be constructed containing cell cycle regulated histone genes under control of a human heat shock gene promoter to allow transcriptional and post-transcriptional regulation to be dissociated. The effects of specific modifications introduced into the histone gene sequence on stability of transcripts during the cell cycle and following inhibition of DNA synthesis will be determined in order to identify sequences involved in regulating mRNA turnover. In addition, equivalent regions of a cell cycle dependent and a cell cycle independent human H4 histone gene will be exchanged. Analysis of the behavior of the resulting chimeric transcripts will indicate the contribution of 5' and 3' domains toward mRNA stability. To gain insight into the mechanism responsible for selective histone mRNA turnover, experiments will be conducted to determine whether the mechanism: 1) operates throughout the cell cycle, 2) is localized to a specific cytoplasmic compartment, 3) is specific for each class of histone mRNA, and 4) involves changes in histone mRNA-protein interactions. Finally, an in vitro system will be developed to allow the assay, identification and characterization of factors mediating histone mRNA turnover. These studies should lead to a better understanding of how histone gene expression is modulated during the cell cycle. Moreover, the results will be relevant to the broader problem of control of mRNA turnover, which is now recognized as an important level of gene regulation during development and in proliferative diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032381-08
Application #
3281171
Study Section
Molecular Biology Study Section (MBY)
Project Start
1983-07-01
Project End
1990-11-30
Budget Start
1990-07-01
Budget End
1990-11-30
Support Year
8
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of South Florida
Department
Type
Schools of Arts and Sciences
DUNS #
City
Tampa
State
FL
Country
United States
Zip Code
33612
Collart, D; Romain, P L; Huebner, K et al. (1992) A human histone H2B.1 variant gene, located on chromosome 1, utilizes alternative 3' end processing. J Cell Biochem 50:374-85
Thor, A; Benz, C; Moore 2nd, D et al. (1991) Stress response protein (srp-27) determination in primary human breast carcinomas: clinical, histologic, and prognostic correlations. J Natl Cancer Inst 83:170-8
Zambetti, G; Ramsey-Ewing, A; Bortell, R et al. (1991) Disruption of the cytoskeleton with cytochalasin D induces c-fos gene expression. Exp Cell Res 192:93-101
Collart, D; Ramsey-Ewing, A; Bortell, R et al. (1991) Isolation and characterization of a cDNA from a human histone H2B gene which is reciprocally expressed in relation to replication-dependent H2B histone genes during HL60 cell differentiation. Biochemistry 30:1610-7
Morris, T D; Weber, L A; Hickey, E et al. (1991) Changes in the stability of a human H3 histone mRNA during the HeLa cell cycle. Mol Cell Biol 11:544-53
Zambetti, G; Fey, E G; Penman, S et al. (1990) Multiple types of mRNA-cytoskeleton interactions. J Cell Biochem 44:177-87
Stein, G S; Lian, J B; Owen, T A (1990) Bone cell differentiation: a functionally coupled relationship between expression of cell-growth- and tissue-specific genes. Curr Opin Cell Biol 2:1018-27
Stein, G S; Lian, J B; Owen, T A (1990) Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation. FASEB J 4:3111-23
Owen, T A; Holthuis, J; Markose, E et al. (1990) Modifications of protein-DNA interactions in the proximal promoter of a cell-growth-regulated histone gene during onset and progression of osteoblast differentiation. Proc Natl Acad Sci U S A 87:5129-33
Zambetti, G; Stein, J; Stein, G (1990) Role of messenger RNA subcellular localization in the posttranscriptional regulation of human histone gene expression. J Cell Physiol 144:175-82

Showing the most recent 10 out of 25 publications