Analysis of Altered Protein Components of Interchromatin Granule Clusters from Breast Cancer Cells
Abstract
Intimate links between the morphology changes of some of sub-nuclear structures and diseases such as cancer have been shown. Thus far, a number of pre-mRNA splicing factors, RNA polymerase II and kinases as well as several cancer related proteins have been reported to be localized to the IGCs. The present study sets a precedent for the global identification of potential differences in the protein composition of IGCs from breast cancer cell nuclei versus normal cells. We have successfully identified the entire protein composition of the isolated IGC fraction from the mouse liver nuclei by proteome analysis using liquid chromatography and tandem mass spectrometry (LC-MS/MS) and combining peptide sequence analysis and database searching. Interestingly, several proteins were identified with well characterized roles in pre-mRNA splicing and yet others known to have structural roles in cells. In addition, we have established the approach to purify IGCs from suspension tissue culture cells, HeLa. The quality of the ICC fraction from the suspension culture has been confirmed using immunoblot analysis. Further, it was observed that the proteins that were enriched in mouse liver IGCs were also found to be enriched in HeLa ICC fractions (splicing factors and RNA processing factors). Work is underway to purify and characterize the IGCs from breast cancer cell lines, HTB 123. Based on these studies it is likely that additional CC proteins will be identified that have an altered organization and/or abundance in breast cancer cells that may lead to altered patterns of gene expression in these cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2002
- Accession Number
- ADA400947
Entities
People
- Prasanth K. Kannanganattu