Genomic and proteomic analysis of the myeloid differentiation program: global analysis of gene expression during induced differentiation in the MPRO cell line
Department of Pediatrics
Animals; Cell Differentiation; Cell Line; Electrophoresis, Gel, Two-Dimensional; Gene Expression Profiling; *Gene Expression Regulation, Developmental; *Genomics; Hematopoietic Stem Cells; Mass Spectrometry; Mice; Myeloid Cells; Myelopoiesis; Neutrophils; Oligonucleotide Array Sequence Analysis; Proteins; *Proteomics; RNA, Messenger; Receptors, Retinoic Acid; Reproducibility of Results; Transcription Factors; Tretinoin
Hematology | Oncology | Pediatrics
We have used an approach using 2-dimensional gel electrophoresis with mass spectrometry analysis combined with oligonucleotide chip hybridization for a comprehensive and quantitative study of the temporal patterns of protein and mRNA expression during myeloid development in the MPRO murine cell line. This global analysis detected 123 known proteins and 29 "new" proteins out of 220 protein spots identified by tandem mass spectroscopy, including proteins in 12 functional categories such as transcription factors and cytokines. Bioinformatic analysis of these proteins revealed clusters with functional importance to myeloid differentiation. Previous analyses have found that for a substantial number of genes the absolute amount of protein in the cell is not strongly correlated to the amount of mRNA. These conclusions were based on simultaneous measurement of mRNA and protein at just a single time point. Here, however, we are able to investigate the relationship between mRNA and protein in terms of simultaneous changes in their levels over multiple time points. This is the first time such a relationship has been studied, and we find that it gives a much stronger correlation, consistent with the hypothesis that a substantial proportion of protein change is a consequence of changed mRNA levels, rather than posttranscriptional effects. Cycloheximide inhibition also showed that most of the proteins detected by gel electrophoresis were relatively stable. Specific investigation of transcription factor mRNA representation showed considerable similarity to those of mature human neutrophils and highlighted several transcription factors and other functional nuclear proteins whose mRNA levels change prominently during MPRO differentiation but which have not been investigated previously in the context of myeloid development. Data are available online at http://bioinfo.mbb.yale.edu/expression/myelopoiesis.
DOI of Published Version
Blood. 2002 Nov 1;100(9):3209-20. doi 10.1182/blood-2002-03-0850
Lian, Zheng; Kluger, Yuval; Greenbaum, Dov S.; Tuck, David; Gerstein, Mark B.; Berliner, Nancy; Weissman, Sherman M.; and Newburger, Peter E., "Genomic and proteomic analysis of the myeloid differentiation program: global analysis of gene expression during induced differentiation in the MPRO cell line" (2002). Hematology/Oncology. 41.