Multiplex selection technique (MuST): An approach to clone transcription factor binding sites

Girish N. Nallur, Kulkarni Prakash, Sherman M. Weissman

Research output: Contribution to journalArticle

Abstract

We have used a multiplex selection approach to construct a library of DNA- protein interaction sites recognized by many of the DNA-binding proteins present in a cell type. An estimated minimum of two-thirds of the binding sites present in a library prepared from activated Jurkat T cells represent authentic transcription factor binding sites. We used the library for isolation of 'optimal' binding site probes that facilitated cloning of a factor and to identify binding activities induced within 2 hr of activation of Jurkat cells. Since a large fraction of the oligonucleotides obtained appear to represent 'optimal' binding sites for sequence-specific DNA- binding proteins, it is feasible to construct a catalog of consensus binding sites for DNA-binding proteins in a given cell type. Qualitative and quantitative comparisons of the catalogs of binding site sequences from various cell types could provide valuable insights into the process of differentiation acting at the level of transcriptional control.

Original languageEnglish (US)
Pages (from-to)1184-1189
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number3
DOIs
StatePublished - Feb 6 1996

Keywords

  • Ets protein
  • T cells
  • cold shock proteins
  • transcription factors

ASJC Scopus subject areas

  • General

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