Abstract

This study will focus on the identification and characterization of nuclear matrix proteins (NMPs) of the rat osteosarcoma (ROS) cell. Our working hypothesis is that NMPs unique to the osteosarcoma cell are related to loss of growth and control and deregulation of differentiation related genes. We have identified several NMPs in the ROS cell line 17/2.8 that are not observed in normal rat osteoblast cells at any stage of differentiation. We will further purify and characterize these proteins and other NMPS, at least 10 fold more abundant in the osteosarcoma cells compared to diploid cells, by a combination of two-dimensional gel electrophoresis, reparative gel electrophoresis, HPLC and other appropriate techniques. Peptide sequences will be determined directly from the purified proteins. Utilizing peptide sequence data for each NMP, we will clone and sequence cDNAs for each protein. These molecular probes will also be used to study expression and regulation the NMPs in several osteosarcoma cell lines. Monoclonal antibodies (produced from either pure protein or from a fusion protein) will be used to characterized the biologically important properties of the expression of each NMP including the nuclear localization, cell cycle alterations and post-transnational modifications that each protein undergoes in the osteosarcoma cell. From an analysis of the amino acid sequence and the biological properties described above, we will begin to address the question of protein function using transfection constructs and antisense down regulation protocols where preliminary evidence indicates that these approaches will permit a meaningful functional analysis. These studies will identify NMPs that contribute to alterations in nuclear architecture and deregulation of growth/differentiation relationships in osteosarcoma cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR045688-01
Application #
2777891
Study Section
Project Start
Project End
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
1
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
University of Massachusetts Medical School Worcester
Department
Type
DUNS #
660735098
City
Worcester
State
MA
Country
United States
Zip Code
01655

  Publications

Ali, Syed A; Zaidi, Sayyed K; Dobson, Jason R et al. (2010) Transcriptional corepressor TLE1 functions with Runx2 in epigenetic repression of ribosomal RNA genes. Proc Natl Acad Sci U S A 107:4165-9
Stein, G S; Stein, J L; Lian, J B et al. (2004) Nuclear microenvironments: an architectural platform for the convergence and integration of transcriptional regulatory signals. Eur J Histochem 48:65-76
Stein, Gary S; Lian, Jane B; van Wijnen, Andre J et al. (2004) Runx2 control of organization, assembly and activity of the regulatory machinery for skeletal gene expression. Oncogene 23:4315-29
Stein, Gary S; Lian, Jane B; Montecino, Martin et al. (2003) Nuclear microenvironments support physiological control of gene expression. Chromosome Res 11:527-36
Lian, Jane B; Stein, Janet L; Stein, Gary S et al. (2003) Runx2/Cbfa1 functions: diverse regulation of gene transcription by chromatin remodeling and co-regulatory protein interactions. Connect Tissue Res 44 Suppl 1:141-8
Stein, Gary S; Lian, Jane B; Stein, Janet L et al. (2003) Temporal and spatial parameters of skeletal gene expression: targeting RUNX factors and their coregulatory proteins to subnuclear domains. Connect Tissue Res 44 Suppl 1:149-53
Stein, Gary S; Lian, Jane B; Stein, Janet L et al. (2003) Intranuclear organization of RUNX transcriptional regulatory machinery in biological control of skeletogenesis and cancer. Blood Cells Mol Dis 30:170-6
Stein, Gary S; Lian, Jane B; Stein, Janet L et al. (2003) Intranuclear trafficking of transcription factors: Requirements for vitamin D-mediated biological control of gene expression. J Cell Biochem 88:340-55
Drissi, Hicham; Pouliot, Arlyssa; Koolloos, Christian et al. (2002) 1,25-(OH)2-vitamin D3 suppresses the bone-related Runx2/Cbfa1 gene promoter. Exp Cell Res 274:323-33
Zaidi, Sayyed K; Sullivan, Andrew J; van Wijnen, Andre J et al. (2002) Integration of Runx and Smad regulatory signals at transcriptionally active subnuclear sites. Proc Natl Acad Sci U S A 99:8048-53

Showing the most recent 10 out of 30 publications