Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Gerhard Mittler
Proteomics and Mass Spectrometry, Max Planck Institute of Immunobiology and Epignetics,
Freiburg
Germany
Research Article
SILAC-Based Quantitative Proteomics Approach to Identify Transcription
Factors Interacting with a Novel Cis-Regulatory Element
Author(s): Ngo Tat Trung, Rudolf Engelke and Gerhard MittlerNgo Tat Trung, Rudolf Engelke and Gerhard Mittler
The motif TMTCGCGANR (M being C or A, R being A or G, and N any nucleotide) called M8 was discovered as a putative cis-regulatory element present in 368 human gene promoters. Of these, 236 (64%) are conserved within promoter sequences of four related organisms: human, mouse, rat and dog. However, transcription factors (TFs) interacting with the M8 motif has not yet been described. We previously reported the use of quantitative proteomics coupled to one-step DNA affinity purification as a means of screening for TFs associated with given functional DNA elements. The procedure is performed in-vitro employing SILAC-labeled nuclear extracts and making use of a well-characterized cis-regulatory motifs. Building on that, in this study we have combined our method with statistical analysis to filter out false positive hits from the one-step DNA affinity pull-down experiments. This resulted in .. View More»
DOI:
10.4172/jpb.1000306