Expression profiling of serum inducible genes identifies a subset of SRF target genes that are MKL dependent

BMC Molecular Biology
Ahalya Selvaraj, R Prywes

Abstract

Serum Response Factor (SRF) is a transcription factor that is required for the expression of many genes including immediate early genes, cytoskeletal genes, and muscle-specific genes. SRF is activated in response to extra-cellular signals by its association with a diverse set of co-activators in different cell types. In the case of the ubiquitously expressed immediate early genes, the two sets of SRF binding proteins that regulate its activity are the TCF family of proteins that include Elk1, SAP1 and SAP2 and the myocardin-related MKL family of proteins that include MKL1 and MKL2 (also known as MAL, MRTF-A and -B and BSAC). In response to serum or growth factors these two classes of co-activators are activated by different upstream signal transduction pathways. However, it is not clear how they differentially activate SRF target genes. In order to identify the serum-inducible SRF target genes that are specifically dependent on the MKL pathway, we have performed microarray experiments using a cell line that expresses dominant negative MKL1. This approach was used to identify SRF target genes whose activation is MKL-dependent. Twenty-eight of 150 serum-inducible genes were found to be MKL-dependent. The promoters of the serum-in...Continue Reading

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Methods Mentioned

BETA
GTPase
immunoprecipitation
gel mobility shift
PCR
chip
immunprecipitations
in vitro transcription
chips

Software Mentioned

dChip
MEME ( Multiple EM for Motif Elicitation )
Affymetrix Microarray Suite
Excel
MEME

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