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A high-throughput method to examine protein-nucleotide interactions identifies targets of the bacterial transcriptional regulatory protein fur. | LitMetric

A high-throughput method to examine protein-nucleotide interactions identifies targets of the bacterial transcriptional regulatory protein fur.

PLoS One

Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston University, Boston, Massachusetts, United States of America; Department of Microbiology, Boston University School of Medicine, Boston University, Boston, Massachusetts, United States of America.

Published: January 2015

AI Article Synopsis

  • The Ferric uptake regulatory protein (Fur) regulates gene transcription in response to iron in pathogenic bacteria like Neisseria gonorrhoeae, which causes gonorrhea.
  • A high-throughput analysis method called Interferometric Reflectance Imaging Sensor (IRIS) was used to study Fur-DNA interactions and demonstrated that 70% of predicted Fur binding sites in iron-induced gene promoters successfully bound to Fur.
  • The study combined binding data with mRNA expression from a gonococcal fur mutant, revealing five new genes that are directly regulated by Fur in Neisseria gonorrhoeae.

Article Abstract

The Ferric uptake regulatory protein (Fur) is a transcriptional regulatory protein that functions to control gene transcription in response to iron in a number of pathogenic bacteria. In this study, we applied a label-free, quantitative and high-throughput analysis method, Interferometric Reflectance Imaging Sensor (IRIS), to rapidly characterize Fur-DNA interactions in vitro with predicted Fur binding sequences in the genome of Neisseria gonorrhoeae, the causative agent of the sexually transmitted disease gonorrhea. IRIS can easily be applied to examine multiple protein-protein, protein-nucleotide and nucleotide-nucleotide complexes simultaneously and demonstrated here that seventy percent of the predicted Fur boxes in promoter regions of iron-induced genes bound to Fur in vitro with a range of affinities as observed using this microarray screening technology. Combining binding data with mRNA expression levels in a gonococcal fur mutant strain allowed us to identify five new gonococcal genes under Fur-mediated direct regulation.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014563PMC
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0096832PLOS

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