Sequence specificity of single-stranded DNA-binding proteins: a novel DNA microarray approach

Hugh P Morgan, Peter Estibeiro, Martin A Wear, Klaas E A Max, Udo Heinemann, Liza Cubeddu, Maurice P Gallagher, Peter J Sadler, Malcolm D Walkinshaw

Research output: Contribution to journalArticlepeer-review

Abstract

We have developed a novel DNA microarray-based approach for identification of the sequence-specificity of single-stranded nucleic-acid-binding proteins (SNABPs). For verification, we have shown that the major cold shock protein (CspB) from Bacillus subtilis binds with high affinity to pyrimidine-rich sequences, with a binding preference for the consensus sequence, 5'-GTCTTTG/T-3'. The sequence was modelled onto the known structure of CspB and a cytosine-binding pocket was identified, which explains the strong preference for a cytosine base at position 3. This microarray method offers a rapid high-throughput approach for determining the specificity and strength of ss DNA-protein interactions. Further screening of this newly emerging family of transcription factors will help provide an insight into their cellular function.
Original languageEnglish
Article numbere75
Number of pages9
JournalNucleic Acids Research
Volume35
Issue number10
DOIs
Publication statusPublished - 2007

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