Abstract

The primary goal of the proposed research is to investigate the biochemical nature and function(s) of mitotic and interphase kinetochore specific chromatin components. The proposed investigations exploit the fact that many scleroderma patients produce high titer antibodies that bind specifically to mammalian and avian kinetochores and interphase kinetochore element antigens. We have recently developed methods for the solubilization of kinetochore specific antigen from isolated nuclei, and for the assay of solubilized antigen during steps of purification. Preliminary experiments suggests kinetochore specific epitopes exist on a subclass of mononucleosomes. The recently developed assay for kinetochore antigen will be used in conjunction with established biochemical and immunologic methods for the fractionation and characterization of chromatin to 1) investigate the biochemical nature of postmitotic chicken erythrocyte kinetochore chromatin, 2) identify kinetochore specific DNA sequences from chicken, and 3) develop monoclonal antibody probes to study the functions of interphase persistant kinetochore components in vivo. This project is relevant to understanding human disorders in which orderly replication, expression and transmittal of genetic information is disturbed; such as aneuploid birth defects and cancer. Further, kinetochore DNA can potentially be used as a vector for introducing stable genetic material into cells for research purposes and therapy of genetic disorders.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032022-03
Application #
3280570
Study Section
Molecular Biology Study Section (MBY)
Project Start
1984-04-01
Project End
1987-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109

  Publications

Palmer, D K; Angello, J C; Margolis, R L (1997) 2-Aminopurine induces spindle cell morphology in MM14 myoblasts in the absence of differentiation signals. Exp Cell Res 230:262-74
Bosc, C; Cronk, J D; Pirollet, F et al. (1996) Cloning, expression, and properties of the microtubule-stabilizing protein STOP. Proc Natl Acad Sci U S A 93:2125-30
Martineau, S N; Andreassen, P R; Margolis, R L (1995) Delay of HeLa cell cleavage into interphase using dihydrocytochalasin B: retention of a postmitotic spindle and telophase disc correlates with synchronous cleavage recovery. J Cell Biol 131:191-205
Andreassen, P R; Margolis, R L (1994) Microtubule dependency of p34cdc2 inactivation and mitotic exit in mammalian cells. J Cell Biol 127:789-802
Cool, D E; Andreassen, P R; Tonks, N K et al. (1992) Cytokinetic failure and asynchronous nuclear division in BHK cells overexpressing a truncated protein-tyrosine-phosphatase. Proc Natl Acad Sci U S A 89:5422-6
Andreassen, P R; Margolis, R L (1992) 2-Aminopurine overrides multiple cell cycle checkpoints in BHK cells. Proc Natl Acad Sci U S A 89:2272-6
Palmer, D K; O'Day, K; Trong, H L et al. (1991) Purification of the centromere-specific protein CENP-A and demonstration that it is a distinctive histone. Proc Natl Acad Sci U S A 88:3734-8
Andreassen, P R; Palmer, D K; Wener, M H et al. (1991) Telophase disc: a new mammalian mitotic organelle that bisects telophase cells with a possible function in cytokinesis. J Cell Sci 99 ( Pt 3):523-34
Andreassen, P R; Margolis, R L (1991) Induction of partial mitosis in BHK cells by 2-aminopurine. J Cell Sci 100 ( Pt 2):299-310
Palmer, D K; O'Day, K; Wener, M H et al. (1987) A 17-kD centromere protein (CENP-A) copurifies with nucleosome core particles and with histones. J Cell Biol 104:805-15

Showing the most recent 10 out of 12 publications